AAK1

Existing Annotations Review

GO Term	Evidence	Action	Reason
GO:0098793 presynapse	IBA GO_REF:0000033	KEEP AS NON CORE	Summary: The IBA places AAK1 at the presynapse from a PANTHER family node and rat Aak1 evidence. This is biologically plausible, but it is a neuronal-context localization rather than the general core function of human AAK1. Reason: AAK1 was reported to be enriched at presynaptic terminals in cultured rat hippocampal neurons (PMID:11877461). Retain the phylogenetic localization as a conserved, context-specific site of action, without treating it as the universal location or defining function of human AAK1. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: PANTHER:PTN000539992 · PANTHER AAK1 family node SOURCE WEAK OR INFERRED The PANTHER node carries the phylogenetic inference and is not an independent localization experiment; it is correlated with the rat donor. RGD:1305520 · rat Aak1 SUPPORTS TRANSFER The rat ortholog is an appropriate donor for the conserved neuronal localization, although presynapse is contextual for the human protein. Supporting Evidence: PMID:11877461 In neuronal cells, AAK1 is enriched at presynaptic terminals
GO:0004674 protein serine/threonine kinase activity	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic assignment of protein serine/threonine kinase activity. This is the defining catalytic function of AAK1 and is independently established by direct biochemical studies. Reason: The IBA is at the appropriate specificity: human AAK1 is a catalytically active NAK-family serine/threonine kinase that phosphorylates AP2M1 and other protein substrates (PMID:11877457, PMID:11877461, PMID:42082516). Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN000539992 · PANTHER AAK1 family node SOURCE WEAK OR INFERRED The family node is the inference carrier and is correlated with the rat donor rather than an independent biochemical observation. RGD:1305520 · rat Aak1 SUPPORTS TRANSFER The orthologous kinase supports transfer, and direct human biochemical evidence independently confirms the target function. Supporting Evidence: PMID:11877457 Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
GO:0035612 AP-2 adaptor complex binding	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic assignment of binding to the AP-2 adaptor complex. Direct work shows AAK1 association with AP2 and interaction with the alpha-adaptin ear, making this a core substrate-recruitment/scaffolding function. Reason: AP-2 binding is directly tied to AAK1 recruitment and AP2M1 phosphorylation during clathrin-mediated endocytosis (PMID:11877461, PMID:12952931, PMID:17494869). The IBA includes the human target itself among its sources, so that item is self-supporting and adds no independent propagation evidence; the annotation remains secure from direct studies. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN000539992 · PANTHER AAK1 family node SOURCE WEAK OR INFERRED This is the phylogenetic inference node, correlated with the donor set. RGD:1305520 · rat Aak1 SUPPORTS TRANSFER The rat ortholog supports conservation of AP-2 association. UniProtKB:Q2M2I8 · human AAK1 target record CIRCULAR OR REDUNDANT The target protein is listed as its own IBA source and therefore cannot serve as independent evidence for propagation. Supporting Evidence: PMID:11877461 Thus, these results demonstrate that AAK1 interacts directly with α-adaptin in vitro.
GO:2000369 regulation of clathrin-dependent endocytosis	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic assignment of regulation of clathrin-dependent endocytosis. This is the central biological role of AAK1 and is supported by multiple direct perturbation and biochemical studies. Reason: AAK1 regulates cargo capture and coated-pit maturation through AP2M1 phosphorylation and has experimentally demonstrated effects on transferrin, LRP, and other cargo internalization (PMID:11877457, PMID:11877461, PMID:12952931, PMID:17494869, PMID:30605688, PMID:34315807). The PMID:12952931 uptake phenotype came from AAK1 overexpression, while about 80% knockdown produced no detectable uptake phenotype, so it is corroborating but not decisive evidence. The human target appears among the IBA sources and is treated as redundant rather than independent evidence. Propagation Review Root cause: NO FAILURE CORE Sources checked: PANTHER:PTN000539992 · PANTHER AAK1 family node SOURCE WEAK OR INFERRED The family node is the correlated inference carrier. RGD:1305520 · rat Aak1 SUPPORTS TRANSFER Rat Aak1 supports conservation of the endocytic regulatory role. UniProtKB:F1SPM8 · pig AAK1 SUPPORTS TRANSFER Pig AAK1 is an orthologous donor supporting the transfer. UniProtKB:Q2M2I8 · human AAK1 target record CIRCULAR OR REDUNDANT Self-support in the IBA source set is redundant and is not counted as an independent line of evidence. Supporting Evidence: PMID:11877461 Together, these results provide strong evidence that AAK1 is the endogenous mu 2 kinase and plays a regulatory role in clathrin-mediated endocytosis.
GO:0004672 protein kinase activity	IEA GO_REF:0000002	MODIFY	Summary: InterPro electronically maps the AAK1 kinase-domain signature to the broad term protein kinase activity. The catalytic class is correct, but a more informative serine/threonine-specific term is already supported directly. Reason: AAK1 is not merely an unspecified protein kinase; biochemical and structural evidence establishes serine/threonine protein kinase activity. Replace the broad parent with GO:0004674, consolidating with existing GO:0004674 rows. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: GRANULARITY MISMATCH Sources checked: InterPro:IPR008271 · protein kinase active-site signature SUPPORTS SOURCE BUT NOT TARGET The signature supports kinase activity but the electronic mapping is broader than the experimentally established serine/threonine specificity; it is also correlated with the other kinase-domain mappings. Proposed replacements: protein serine/threonine kinase activity Supporting Evidence: PMID:11877461 Here we identify and characterize a novel member of the Prk/Ark family of serine/threonine kinases, adaptor-associated kinase (AAK)1.
GO:0004674 protein serine/threonine kinase activity	IEA GO_REF:0000003	ACCEPT	Summary: EC-to-GO electronic assignment of protein serine/threonine kinase activity. It matches AAK1's experimentally established catalytic class. Reason: EC 2.7.11.1 is correctly mapped to the core molecular function, which is independently supported by direct AP2M1 phosphorylation and human AAK1 kinase structures. The EC mapping is corroborative and correlated with the other electronic kinase-domain rows, not a separate experiment. Propagation Review Root cause: NO FAILURE CORE Sources checked: EC:2.7.11.1 · non-specific serine/threonine protein kinase EC class SUPPORTS TRANSFER The EC mapping has the correct catalytic scope; it is an electronic classification correlated with the InterPro and Rhea mappings.
GO:0005524 ATP binding	IEA GO_REF:0000002	KEEP AS NON CORE	Summary: InterPro kinase-domain mapping to ATP binding. ATP binding is mechanistically correct for AAK1 but is generic relative to its informative catalytic activity. Reason: The AAK1 kinase domain contains the conserved ATP-binding pocket and direct structural work resolves inhibitor binding in that pocket (PMID:26853940). Retain the annotation as a valid cofactor-binding property, but do not elevate generic ATP binding to a core functional summary. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: InterPro:IPR000719 · protein kinase domain SUPPORTS TRANSFER The domain supports ATP binding, but this electronic row is correlated with the other kinase-domain mappings and adds no independent experiment. Supporting Evidence: PMID:26853940 Here we report the first high-resolution structures of kinases AAK1 and BIKE in complex with two drug candidates.
GO:0005886 plasma membrane	IEA GO_REF:0000044	ACCEPT	Summary: Electronic localization to the plasma membrane derived from the UniProt subcellular-location vocabulary. This is consistent with AAK1 action at plasma-membrane clathrin-coated pits and with direct human-cell imaging. Reason: AAK1 acts peripherally at the plasma membrane during cargo internalization. Direct RPE-cell imaging found broad plasma-membrane AAK1 distribution (PMID:42082516), independently supporting the location. The vocabulary mapping is generated from the same UniProt target record and is therefore correlated, not an independent localization experiment. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB-SubCell:SL-0039 · UniProt cell membrane vocabulary term CIRCULAR OR REDUNDANT This IEA maps the target record's own subcellular-location statement; it is valid but self-derived and correlated with other UniProt location rows. Supporting Evidence: PMID:42082516 In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
GO:0005905 clathrin-coated pit	IEA GO_REF:0000044	ACCEPT	Summary: Electronic localization to clathrin-coated pits from the UniProt subcellular-location vocabulary. This is a principal site at which AAK1 controls AP2 and cargo capture. Reason: Direct localization and biochemical work place AAK1 with AP2 and clathrin at coated pits (PMID:11877461), while clathrin activates AAK1 (PMID:14617351). The mapping is correct but derives from the same UniProt record, so it should not be counted as independent evidence. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB-SubCell:SL-0069 · UniProt clathrin-coated pit vocabulary term CIRCULAR OR REDUNDANT This is a self-derived vocabulary mapping from the AAK1 UniProt record, correlated with its other mapped localization rows. Supporting Evidence: PMID:11877461 AAK1 also colocalized to endocytic structures in nonneuronal cells.
GO:0098793 presynapse	IEA GO_REF:0000044	KEEP AS NON CORE	Summary: Electronic presynapse localization mapped from the UniProt record. The location is plausible from neuronal ortholog evidence but is tissue-specific. Reason: Rat-neuron experiments reported enrichment at presynaptic terminals (PMID:11877461). Retain this as contextual neuronal localization rather than a general core location of human AAK1. Because the IEA is mapped from the target record's own location statement, it is correlated/self-derived. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: UniProtKB-SubCell:SL-0516 · UniProt presynapse vocabulary term CIRCULAR OR REDUNDANT The source is the target record's own vocabulary assertion and is not an independent human presynaptic localization experiment. Supporting Evidence: PMID:11877461 In neuronal cells, AAK1 is enriched at presynaptic terminals
GO:0106310 protein serine kinase activity	IEA GO_REF:0000116	ACCEPT	Summary: Rhea electronically maps the serine-specific protein phosphorylation reaction to the current protein serine kinase activity term. This is one valid subactivity within AAK1's broader serine/threonine kinase activity. Reason: RHEA:17989 directly supports the serine-specific electronic mapping, and this annotation can coexist with the broader GO:0004674 rows. AAK1 also has strong threonine-substrate evidence, but current GO has no separate primary protein threonine kinase term: GO:0106311 is an alternate ID of GO:0004674, not a usable sibling term. Retain the valid serine subactivity rather than generalizing it or writing an alternate identifier. Propagation Review Root cause: NO FAILURE CORE Sources checked: RHEA:17989 · L-seryl-protein phosphorylation reaction SUPPORTS TRANSFER The reaction directly supports the serine subactivity. It is correlated with the UniProt catalytic record and does not negate AAK1's independently supported threonine activity.
GO:0005515 protein binding	IPI PMID:22645275 Identification of novel ATP13A2 interactors and their role i...	MARK AS OVER ANNOTATED	Summary: GOA/IntAct records AAK1 as an ATP13A2 interactor from a yeast two-hybrid screen. The cached abstract describes the 43-interactor screen but does not name AAK1, and the full text is unavailable locally, so the specific pair cannot be independently verified here. Reason: Retain and defer to the experimental interaction provenance; this action does not reject or question the AAK1-ATP13A2 pair. It concerns only GO:0005515, which is an uninformative generic molecular-function term and does not capture the interaction partner, mechanism, or biological role. Mark the term as over-annotated while recording that the pair-level evidence remains unverified from the locally accessible abstract. Supporting Evidence: PMID:22645275 Utilizing a split-ubiquitin membrane yeast two-hybrid system that was developed to identify interacting partners of full-length integral membrane proteins, we identified 43 novel interactors
GO:0061024 membrane organization	TAS Reactome:R-HSA-199991	KEEP AS NON CORE	Summary: Reactome assigns AAK1 to the broad membrane-trafficking umbrella and maps that participation to membrane organization. This captures pathway context but is broader than AAK1's specific clathrin-mediated endocytic role. Reason: Membrane organization is defensible as a high-level consequence of AAK1's endocytic function, but the core role is regulation of clathrin-dependent endocytosis through AP2M1 phosphorylation. Retain this TAS annotation as broad pathway context rather than as a defining function.
GO:0004674 protein serine/threonine kinase activity	TAS Reactome:R-HSA-8856813	ACCEPT	Summary: Reactome represents AAK1 phosphorylating the AP-2 mu subunit and maps the event to protein serine/threonine kinase activity. This matches the core catalytic function established experimentally. Reason: The pathway event is consistent with direct biochemical evidence that AAK1 phosphorylates AP2M1 at Thr-156 (PMID:11877457). Although the TAS is model based, its catalytic assertion is correctly scoped and independently supported. Supporting Evidence: PMID:11877457 Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
GO:0005886 plasma membrane	IDA GO_REF:0000052	ACCEPT	Summary: Human Protein Atlas immunofluorescence places AAK1 at the plasma membrane. This agrees with its role at plasma-membrane clathrin-coated pits. Reason: The direct human-cell localization is biologically consistent with AAK1 cargo internalization and with independent live-cell evidence of plasma-membrane distribution (PMID:42082516). Retain the experimentally assigned location. Supporting Evidence: PMID:42082516 In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
GO:0005886 plasma membrane	ISS GO_REF:0000024	ACCEPT	Summary: Plasma-membrane localization transferred by curator-reviewed similarity from bovine AAK1. This is conserved and independently supported in human cells. Reason: The bovine ortholog is an appropriate donor and direct imaging of human AAK1 supports plasma-membrane distribution (PMID:42082516). The same donor is reused for multiple location rows, so those ISS annotations are correlated rather than independent observations. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:F1MH24 · bovine AAK1 SUPPORTS TRANSFER The ortholog supports transfer; reuse of this single donor for plasma membrane and coated pit makes the inferred rows correlated. Supporting Evidence: PMID:42082516 In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
GO:0098793 presynapse	ISS GO_REF:0000024	KEEP AS NON CORE	Summary: Presynapse localization transferred from rat Aak1. The inference is plausible and supported in rat neurons, but it is a neuronal-context location. Reason: Preserve the ortholog transfer because rat-neuron experiments place AAK1 at presynaptic terminals (PMID:11877461), while recognizing that the human row is model-derived and should not define AAK1's general cellular location. This and other P0C1X8 transfers reuse one donor and are correlated. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: UniProtKB:P0C1X8 · rat Aak1 SUPPORTS TRANSFER Rat Aak1 supports the neuronal localization; reuse of this donor across kinase and neuronal-location rows does not constitute independent evidence. Supporting Evidence: PMID:11877461 In neuronal cells, AAK1 is enriched at presynaptic terminals
GO:0005905 clathrin-coated pit	ISS GO_REF:0000024	ACCEPT	Summary: Clathrin-coated-pit localization transferred from bovine AAK1. The location is central to AAK1's endocytic role and is independently observed in human cells. Reason: AAK1 colocalizes with AP2 and clathrin at coated-pit structures and is activated by assembled clathrin (PMID:11877461, PMID:14617351). The bovine donor is sound, but its reuse with the plasma-membrane ISS row makes the evidence correlated. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:F1MH24 · bovine AAK1 SUPPORTS TRANSFER Bovine AAK1 is a suitable orthologous donor; this is the same donor used for the plasma-membrane transfer and is not an independent second source. Supporting Evidence: PMID:11877461 AAK1 also colocalized to endocytic structures in nonneuronal cells.
GO:0005829 cytosol	TAS Reactome:R-HSA-8856808	KEEP AS NON CORE	Summary: The Reactome AP-2/clathrin recruitment event models AAK1 as cytosolic before or during recruitment to the endocytic machinery. Reason: A cytosolic pool is plausible for this peripheral-membrane kinase, but this TAS is generated from a shared Reactome clathrin-mediated-endocytosis model. The 17 Reactome cytosol rows are correlated pathway-context assertions, not independent localization experiments; retain each as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8856813	KEEP AS NON CORE	Summary: The Reactome AP2M1 phosphorylation event assigns its AAK1 participant to the cytosol. Reason: Cytosolic availability is compatible with recruitment of AAK1 to coated pits, but the location is model context rather than a direct assay. It is correlated with the other 16 cytosol tuples emitted from the same Reactome pathway and is retained as non-core.
GO:0005829 cytosol	TAS Reactome:R-HSA-8866283	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled recruitment of GPCRs to clathrin-coated pits. Reason: This is a correlated participant-location assertion from the shared Reactome endocytosis model, not an independent localization experiment or a distinct AAK1 function. Retain it as non-core pathway context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8867754	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the modeled F-/N-BAR protein recruitment phase of coated-pit maturation. Reason: The cytosolic location is biologically compatible with a recruitable peripheral kinase, but this tuple is one of 17 correlated annotations generated from the same pathway model. Keep it as non-core context rather than independent support.
GO:0005829 cytosol	TAS Reactome:R-HSA-8867756	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in a modeled nascent coated-pit cargo/CLASP recruitment event. Reason: This pathway-derived location is plausible but does not establish a separate localization result. It is correlated with the other Reactome cytosol rows and is retained as non-core endocytic context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868071	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the clathrin-dependent PIK3C2A recruitment event. Reason: The TAS reflects a shared pathway compartment rather than an AAK1-specific localization experiment. It is correlated with the 16 other Reactome cytosol rows and is retained only as non-core pathway context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868072	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled PIK3C2A phosphoinositide phosphorylation stage of coated-pit maturation. Reason: This is a pathway-participant compartment assignment, not direct evidence that independently localizes AAK1. Treat it as correlated with the other Reactome cytosol tuples and retain it as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868230	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled SNX9/actin-machinery recruitment stage of clathrin-mediated endocytosis. Reason: The location derives from the common pathway model and is not an independent AAK1 localization assay. It is correlated with the other 16 cytosol rows and should remain non-core.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868236	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in a modeled BAR-protein/dynamin recruitment event. Reason: Cytosol is plausible for the unbound pool, but this TAS is a correlated pathway-context assertion rather than direct localization evidence. Retain it as non-core.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868648	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled synaptojanin lipid-hydrolysis stage of coated-vesicle formation. Reason: This tuple inherits the pathway compartment and is not independent evidence for AAK1 localization or synaptojanin regulation. It is correlated with the other Reactome cytosol rows and is retained as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868651	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the modeled endophilin/synaptojanin recruitment event. Reason: The assignment is a shared pathway-location statement, not an AAK1-specific experiment. It is correlated with the other 16 Reactome cytosol annotations and is kept only as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868658	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled HSPA8-driven vesicle-uncoating event. Reason: This is correlated compartment context from a broader endocytosis model, not evidence that AAK1 directly performs or controls HSPA8 ATP hydrolysis. Retain the cytosol tuple as non-core.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868659	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the modeled recruitment of auxilins to clathrin-coated vesicles. Reason: The TAS derives from the shared pathway compartment rather than a direct AAK1 localization assay. It is correlated with the other Reactome cytosol tuples and is retained as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868660	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled auxilin/HSPA8 recruitment event. Reason: This participant-location row is correlated with the common Reactome pathway model and does not provide an independent localization observation. Retain it as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8868661	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the modeled dynamin-mediated vesicle scission event. Reason: The tuple inherits a pathway compartment and should not be interpreted as separate evidence that AAK1 controls dynamin catalysis. It is correlated with the other 16 cytosol annotations and is retained as non-core context.
GO:0005829 cytosol	TAS Reactome:R-HSA-8871193	KEEP AS NON CORE	Summary: Reactome assigns cytosolic AAK1 in the modeled dissociation of AAK1 and dephosphorylation of AP2M1. Reason: Cytosolic release after coated-pit action is mechanistically plausible, but this is model-derived and correlated with the other Reactome cytosol rows. It is retained as non-core rather than treated as direct localization evidence.
GO:0005829 cytosol	TAS Reactome:R-HSA-8871194	KEEP AS NON CORE	Summary: Reactome places AAK1 in the cytosol in the modeled RAB5/GAPVD1 binding phase of the AP-2 endocytic pathway. Reason: The TAS is a correlated pathway-participant compartment assignment, not a distinct AAK1 localization experiment. Retain it as non-core context with the other Reactome cytosol tuples.
GO:0004674 protein serine/threonine kinase activity	ISS GO_REF:0000024	ACCEPT	Summary: Protein serine/threonine kinase activity transferred from rat Aak1 by curator-reviewed similarity. This is the defining catalytic activity and is independently established for human AAK1. Reason: The rat ortholog is an appropriate donor, and direct biochemical and structural studies of human AAK1 confirm an active serine/threonine kinase (PMID:11877457, PMID:11877461, PMID:26853940). The same donor is reused for several localization rows, so those transfers are correlated rather than independent support. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:P0C1X8 · rat Aak1 SUPPORTS TRANSFER Rat Aak1 is a suitable orthologous donor; reuse of this one donor across kinase, presynapse, leading-edge, and bouton rows is correlated evidence. Supporting Evidence: PMID:11877457 Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
GO:0005112 Notch binding	IDA PMID:21464124 The adaptor-associated kinase 1, AAK1, is a positive regulat...	KEEP AS NON CORE	Summary: AAK1 directly binds the membrane-tethered activated form of NOTCH1, but not the inactive full-length receptor, in a ligand-activation/endocytic context. Reason: The IDA is directly supported and more informative than generic protein binding. Retain it as a context-specific interaction that helps route activated NOTCH1, rather than treating Notch binding as the defining molecular function of AAK1. Supporting Evidence: PMID:21464124 We show here that AAK1, the adaptor-associated kinase 1, directly interacts with the membrane-tethered active form of Notch released by metalloprotease cleavage.
GO:0006468 protein phosphorylation	IDA PMID:17494869 A novel AAK1 splice variant functions at multiple steps of t...	MODIFY	Summary: AAK1L and AAK1S were tested in kinase assays against the AP-2 mu2 subunit, whose established AAK1 phosphosite is Thr-156. Reason: The study directly assayed both human splice forms and found basal and clathrin-stimulated AP2M1 kinase activity. Replace the broad process term with GO:0018107 peptidyl-threonine phosphorylation, supported by direct mapping of the AAK1-dependent AP2M1 site to Thr-156 in PMID:11877457. Proposed replacements: peptidyl-threonine phosphorylation Supporting Evidence: PMID:17494869 However, in vitro kinase assays reveal little difference between AAK1 isoforms in their basal or clathrin-stimulated kinase activity toward the AP-2 micro2 subunit. PMID:11877457 Phosphorylation of mu 2 occurs at a single threonine residue (Thr-156) and is mediated by the newly discovered adaptor-associated kinase, AAK1, which copurifies with AP2.
GO:0006468 protein phosphorylation	IDA PMID:18657069 AAK1 regulates Numb function at an early step in clathrin-me...	MODIFY	Summary: The source reports AAK1-dependent phosphorylation of NUMB and identifies T102 as the relevant site. A 2026 biochemical study did not detect phosphorylation of a NUMB T102 peptide, creating substrate-site-specific counterevidence. Reason: Replace the broad process term with GO:0018107 peptidyl-threonine phosphorylation. The original study reports a threonine phosphosite, while AP2M1 T156 and newer PDLIM5/Talin1 sites independently establish AAK1-mediated threonine phosphorylation. This replacement does not endorse NUMB T102 as a physiological substrate: that specific assignment is disputed by PMID:42082516. Proposed replacements: peptidyl-threonine phosphorylation Supporting Evidence: PMID:18657069 In this study, we show that Numb binds to and is phosphorylated by adaptor-associated kinase 1 (AAK1), a key endocytic kinase. PMID:42082516 This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
GO:0030136 clathrin-coated vesicle	ISS GO_REF:0000024	ACCEPT	Summary: Clathrin-coated-vesicle localization transferred from pig AAK1. This is consistent with AAK1 association with AP2, clathrin, and isolated coated vesicles during its core endocytic function. Reason: The pig ortholog is an appropriate donor, and biochemical fractionation of mammalian tissues independently recovered AAK1 with clathrin-coated vesicles (PMID:11877461). This donor also supports an IBA process row, making those propagated assertions correlated rather than independent. Propagation Review Root cause: NO FAILURE CORE Sources checked: UniProtKB:F1SPM8 · pig AAK1 SUPPORTS TRANSFER Pig AAK1 supports the conserved coated-vesicle location; reuse of this donor in the IBA endocytosis row is correlated evidence. Supporting Evidence: PMID:11877461 AAK1 is found associated with isolated APs (lane 1), clathrin (lane 2), and CCVs (lane 3), and CCVs from rat liver (lane 4).
GO:0031252 cell leading edge	ISS GO_REF:0000024	KEEP AS NON CORE	Summary: Cell-leading-edge localization transferred from rat Aak1. Direct human-cell work supports leading-edge/endocytic and focal-adhesion recruitment during migration, but this is context-dependent. Reason: Human HeLa imaging reported AAK1 enrichment at the leading edge (PMID:11877461), and newer RPE-cell work links AAK1 recruitment to focal adhesion turnover and migration (PMID:42082516). Retain this localization as a non-core migratory-cell context. The rat donor is reused across several ISS rows and thus does not create independent evidence for each. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: UniProtKB:P0C1X8 · rat Aak1 SUPPORTS TRANSFER Rat Aak1 is an appropriate donor, but this single donor is shared with the kinase, presynapse, and terminal-bouton ISS rows. Supporting Evidence: PMID:11877461 Interestingly, in migrating cells AAK1 was highly enriched at the leading edge
GO:0032880 regulation of protein localization	IDA PMID:18657069 AAK1 regulates Numb function at an early step in clathrin-me...	KEEP AS NON CORE	Summary: AAK1 perturbation changes NUMB distribution between the plasma membrane and perinuclear endosomes, directly supporting regulation of protein localization. Reason: Preserve the reported localization phenotype even though the proposed direct phosphorylation of NUMB T102 is challenged by PMID:42082516. The redistribution result supports a real, substrate/context-specific regulatory output, but it is secondary to AAK1's core AP2/endocytic function. Supporting Evidence: PMID:18657069 We find that AAK1 redistributes Numb to perinuclear endosomes when overexpressed, while kinase depletion causes Numb to accumulate at the plasma membrane. PMID:42082516 This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
GO:0035612 AP-2 adaptor complex binding	IDA PMID:17494869 A novel AAK1 splice variant functions at multiple steps of t...	ACCEPT	Summary: The source supports association of AAK1 isoforms with the AP-2 machinery and assays AP2M1 as their kinase substrate. Reason: Although the locally cached source exposes only the abstract and does not spell out every binding assay, the experimental curator had access to the full study. Direct AP-2/alpha-adaptin binding was independently demonstrated in PMID:11877461, so the informative AP-2-complex-binding term is retained. Supporting Evidence: PMID:11877461 Thus, these results demonstrate that AAK1 interacts directly with α-adaptin in vitro.
GO:0043195 terminal bouton	ISS GO_REF:0000024	KEEP AS NON CORE	Summary: Terminal-bouton localization transferred from rat Aak1. It is compatible with reported presynaptic-terminal enrichment but is a neuronal model-derived sublocation rather than a general human AAK1 location. Reason: Keep the ortholog transfer as contextual neuronal localization. The cached experimental evidence supports presynaptic-terminal enrichment (PMID:11877461) but not a uniquely required terminal-bouton role, and the same rat donor is reused across multiple ISS rows. Propagation Review Root cause: NO FAILURE NON CORE Sources checked: UniProtKB:P0C1X8 · rat Aak1 SUPPORTS TRANSFER The donor supports neuronal terminal localization, but this is the same source reused for presynapse, kinase, and leading-edge transfers. Supporting Evidence: PMID:11877461 In neuronal cells, AAK1 is enriched at presynaptic terminals
GO:0045747 positive regulation of Notch signaling pathway	IDA PMID:21464124 The adaptor-associated kinase 1, AAK1, is a positive regulat...	KEEP AS NON CORE	Summary: AAK1 promotes activated NOTCH1 routing and transcriptional output upstream of gamma-secretase, supporting positive regulation of Notch signaling in the tested ligand-activation context. Reason: The direct perturbation evidence supports a positive effect on mammalian Notch signaling. Retain it as a context-specific signaling consequence of AAK1's endocytic/adaptor activity, rather than as the kinase's universal core process. Supporting Evidence: PMID:21464124 Moreover, shRNA-mediated AAK1 depletion decreases Notch transcriptional activity in coculture experiments.
GO:0050821 protein stabilization	IDA PMID:21464124 The adaptor-associated kinase 1, AAK1, is a positive regulat...	KEEP AS NON CORE	Summary: AAK1 stabilizes the membrane-tethered activated NOTCH1 fragment and its monoubiquitinated counterpart upstream of gamma-secretase cleavage. Reason: The IDA is supported for the specific activated-NOTCH1 experimental context. Protein stabilization is a secondary, substrate-specific output of AAK1 adaptor/endocytic activity and should not be treated as its defining process. Supporting Evidence: PMID:21464124 Active AAK1 acts upstream of the γ-secretase cleavage by stabilizing both the membrane-tethered activated form of Notch and its monoubiquitinated counterpart.
GO:2000369 regulation of clathrin-dependent endocytosis	IMP PMID:17494869 A novel AAK1 splice variant functions at multiple steps of t...	ACCEPT	Summary: AAK1L-domain overexpression and AAK1 depletion perturb transferrin internalization and recycling, directly demonstrating regulation at multiple steps of the clathrin-dependent endocytic pathway. Reason: This IMP annotation captures the central biological role established by perturbation of AAK1 constructs and abundance. Cargo and direction of effect vary with experimental context, so the sign-neutral regulatory GO term is appropriately scoped. Supporting Evidence: PMID:17494869 These observations suggest that AAK1 functions at multiple steps of the endosomal pathway by regulating transferrin internalization and its rapid recycling back to the plasma membrane from early/sorting endosome.
GO:2000369 regulation of clathrin-dependent endocytosis	IDA PMID:18657069 AAK1 regulates Numb function at an early step in clathrin-me...	ACCEPT	Summary: AAK1 perturbation and NUMB localization/mutant experiments support regulation of cargo-selective clathrin-dependent endocytosis and coated-pit maturation. Reason: Preserve the broad endocytic-regulation annotation: the reported NUMB redistribution and transferrin/LDL uptake phenotypes support it, and AAK1's endocytic role is independently established (PMID:11877461, PMID:17494869). PMID:42082516 disputes direct phosphorylation of the proposed NUMB T102 peptide, so that specific mechanism is treated cautiously without discarding the phenotype or the broader process. Supporting Evidence: PMID:18657069 Overexpression of a Numb point mutant (T102A) that lacks the AAK1 phosphorylation site potently disrupts transferrin and low-density lipoprotein internalization but does not impact EGF uptake. PMID:42082516 This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
GO:0030276 clathrin binding	IDA Q2M2I8-1 PMID:17494869 A novel AAK1 splice variant functions at multiple steps of t...	NEW	Summary: NEW annotation. The long AAK1 isoform contains a second C-terminal clathrin-binding region whose direct interaction with clathrin was tested in protein-interaction assays. Reason: Clathrin binding is an informative molecular function that helps recruit and stimulate AAK1 in the coated-pit machinery. The isoform field records that the tested CBD2 construct came from long isoform AAK1L/Q2M2I8-1; it does not claim that clathrin binding is unique to this isoform. Supporting Evidence: PMID:17494869 Protein interaction studies demonstrate that AAK1L CBD2 directly binds clathrin.
GO:2001137 positive regulation of endocytic recycling	IMP Q2M2I8-1 PMID:17494869 A novel AAK1 splice variant functions at multiple steps of t...	NEW	Summary: NEW annotation. AAK1 depletion or interference with its second clathrin-binding region impairs transferrin recycling from early/sorting endosomes, and independent depletion experiments delay short-loop beta3 integrin recycling to the cell surface. Reason: Loss-of-function evidence for two cargos supports a positive contribution to endocytic recycling, a process not captured by the existing annotation to regulation of clathrin-dependent endocytosis. The isoform field records the tested long AAK1L/Q2M2I8-1 constructs without claiming that the shorter isoform cannot contribute. Supporting Evidence: PMID:17494869 Surprisingly, CBD2 overexpression or AAK1 depletion by RNA interference significantly impairs transferrin recycling from the early/sorting endosome. PMID:23781025 siRNA-mediated depletion of either factor delays short-loop β3 integrin recycling from the early endosome back to the cell surface.
GO:0005925 focal adhesion	IDA PMID:42082516 AAK1-mediated phosphorylation of PDLIM5 and Talin1 promotes ...	NEW	Summary: NEW annotation. Live-cell imaging places AAK1 at mature, tension-bearing focal adhesions in collagen-adherent human RPE cells. Reason: The ECM-dependent localization was supported by paxillin colocalization and multiple illumination, membrane, and attachment controls. This is the site of AAK1's directly demonstrated adhesion-turnover activity. Supporting Evidence: PMID:42082516 Thus, AAK1 is specifically recruited to mature, tension-bearing FAs and thereby positioned to efficiently phosphorylate substrates PDLIM5 and Talin1.
GO:0120183 positive regulation of focal adhesion disassembly	IMP PMID:42082516 AAK1-mediated phosphorylation of PDLIM5 and Talin1 promotes ...	NEW	Summary: NEW annotation. AAK1 deletion increases focal-adhesion density and lifetime and slows paxillin exchange, while kinase- and phosphosite-dependent analyses support accelerated release of focal-adhesion components. Reason: Human-cell loss-of-function, live imaging, rescue, and phosphomutant evidence directly supports a positive regulatory role in focal-adhesion disassembly. This is a core AAK1 function distinct from its cargo-trafficking role. Supporting Evidence: PMID:42082516 Because rapid paxillin turnover is essential for efficient cell migration, these data position AAK1 as a positive regulator of FA disassembly and motility.
GO:0030335 positive regulation of cell migration	IMP PMID:42082516 AAK1-mediated phosphorylation of PDLIM5 and Talin1 promotes ...	NEW	Summary: NEW annotation. AAK1 knockout or knockdown slows human RPE-cell migration, with corroborating knockdown phenotypes in SH-SY5Y and MDA-MB-231 cells. Reason: Genetic loss and kinase-inhibitor phenotypes establish a positive, kinase-dependent contribution to cell migration. BMP2K loss did not reproduce the RPE phenotype, supporting an AAK1-specific core role. Supporting Evidence: PMID:42082516 AAK1 deletion or siRNA-mediated knockdown reproducibly impaired RPE migration by ~25% (Fig. 3d, e and Supplementary Fig. 1h), whereas BMP2K loss had no measurable effect, revealing a specific and non-redundant role for AAK1 in cell motility.
GO:0090090 negative regulation of canonical Wnt signaling pathway	IMP PMID:30605688 WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated E...	NEW	Summary: NEW annotation. AAK1 loss or inhibition increases canonical WNT output in multiple human cell models, while AAK1 promotes LRP6 endocytosis and lysosomal degradation. Reason: Perturbation and mechanistic evidence support negative regulation of canonical WNT signaling. This is retained as a context- and cargo-specific output of the endocytic machinery, not as a universal core function of AAK1. Supporting Evidence: PMID:30605688 Together, these results suggest that AAK1 inhibits WNT signaling by inducing CME endocytosis and lysosomal degradation of LRP6.
GO:0160020 positive regulation of ferroptosis	IMP PMID:41407700 AAK1 activation-mediated iron trafficking drives ferroptotic...	NEW	Summary: NEW annotation. In induced ferroptosis models, AAK1 loss reduces lipid peroxidation and ferroptotic death, whereas activated AAK1 drives AP2M1-dependent TFR1 internalization and iron accumulation. Reason: Genetic and mechanistic evidence supports positive regulation of ferroptosis in the tested cancer-cell and xenograft settings. This stimulus-dependent, cargo-specific role is non-core rather than a constitutive output of AAK1. Supporting Evidence: PMID:41407700 Mechanistically, activated AAK1 phosphorylates AP2M1, which facilitates the recruitment of clathrin to mediate the endocytosis of TFR1, increasing the levels of both cellular total iron and ferrous iron and thereby promoting ferroptosis.

📄 View Raw YAML

id: Q2M2I8
gene_symbol: AAK1
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: >-
  AAK1 is a cytosolic, peripheral-membrane serine/threonine kinase and endocytic
  scaffold of the Numb-associated kinase family. It is recruited through AP-2 and
  clathrin interactions and phosphorylates AP2M1/μ2 at Thr-156, increasing AP-2
  affinity for cargo sorting motifs; assembled clathrin stimulates this activity. The
  long AAK1L isoform has an additional C-terminal clathrin-binding region and contributes
  to both cargo internalization and rapid recycling from early/sorting endosomes. Effects
  on uptake depend on cargo and cellular context and overlap with other NAK-family
  kinases. AAK1 also localizes to focal adhesions, where its kinase activity promotes
  adhesion disassembly and cell migration; PDLIM5 Thr-290 and talin-1 Thr-2270 are
  direct in-vitro AAK1/BMP2K substrates implicated in this output. More
  context-specific outputs include activated NOTCH1 stabilization and endosomal
  routing, WNT-LRP6 internalization, and TFR1-dependent iron uptake during ferroptosis.
alternative_products:
- name: 1 (AAK1L {ECO:0000303|PubMed:17494869})
  id: Q2M2I8-1
- name: 2 (AAK1S {ECO:0000303|PubMed:17494869})
  id: Q2M2I8-2
  sequence_note: VSP_039459
existing_annotations:
- term:
    id: GO:0098793
    label: presynapse
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: is_active_in
  supporting_entities:
  - PANTHER:PTN000539992
  - RGD:1305520
  review:
    summary: >-
      The IBA places AAK1 at the presynapse from a PANTHER family node and rat
      Aak1 evidence. This is biologically plausible, but it is a neuronal-context
      localization rather than the general core function of human AAK1.
    action: KEEP_AS_NON_CORE
    reason: >-
      AAK1 was reported to be enriched at presynaptic terminals in cultured rat
      hippocampal neurons (PMID:11877461). Retain the phylogenetic localization as
      a conserved, context-specific site of action, without treating it as the
      universal location or defining function of human AAK1.
    additional_reference_ids:
    - PMID:11877461
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: PANTHER:PTN000539992
        source_label: PANTHER AAK1 family node
        source_status: SOURCE_WEAK_OR_INFERRED
        comment: >-
          The PANTHER node carries the phylogenetic inference and is not an
          independent localization experiment; it is correlated with the rat donor.
      - source_id: RGD:1305520
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The rat ortholog is an appropriate donor for the conserved neuronal
          localization, although presynapse is contextual for the human protein.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: In neuronal cells, AAK1 is enriched at presynaptic terminals
- term:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  supporting_entities:
  - PANTHER:PTN000539992
  - RGD:1305520
  review:
    summary: >-
      Phylogenetic assignment of protein serine/threonine kinase activity. This is
      the defining catalytic function of AAK1 and is independently established by
      direct biochemical studies.
    action: ACCEPT
    reason: >-
      The IBA is at the appropriate specificity: human AAK1 is a catalytically
      active NAK-family serine/threonine kinase that phosphorylates AP2M1 and other
      protein substrates (PMID:11877457, PMID:11877461, PMID:42082516).
    additional_reference_ids:
    - PMID:11877457
    - PMID:11877461
    - PMID:42082516
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: PANTHER:PTN000539992
        source_label: PANTHER AAK1 family node
        source_status: SOURCE_WEAK_OR_INFERRED
        comment: >-
          The family node is the inference carrier and is correlated with the rat
          donor rather than an independent biochemical observation.
      - source_id: RGD:1305520
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The orthologous kinase supports transfer, and direct human biochemical
          evidence independently confirms the target function.
    supported_by:
    - reference_id: PMID:11877457
      supporting_text: >-
        Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
- term:
    id: GO:0035612
    label: AP-2 adaptor complex binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  supporting_entities:
  - PANTHER:PTN000539992
  - RGD:1305520
  - UniProtKB:Q2M2I8
  review:
    summary: >-
      Phylogenetic assignment of binding to the AP-2 adaptor complex. Direct work
      shows AAK1 association with AP2 and interaction with the alpha-adaptin ear,
      making this a core substrate-recruitment/scaffolding function.
    action: ACCEPT
    reason: >-
      AP-2 binding is directly tied to AAK1 recruitment and AP2M1 phosphorylation
      during clathrin-mediated endocytosis (PMID:11877461, PMID:12952931,
      PMID:17494869). The IBA includes the human target itself among its sources, so
      that item is self-supporting and adds no independent propagation evidence;
      the annotation remains secure from direct studies.
    additional_reference_ids:
    - PMID:11877461
    - PMID:12952931
    - PMID:17494869
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: PANTHER:PTN000539992
        source_label: PANTHER AAK1 family node
        source_status: SOURCE_WEAK_OR_INFERRED
        comment: >-
          This is the phylogenetic inference node, correlated with the donor set.
      - source_id: RGD:1305520
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: The rat ortholog supports conservation of AP-2 association.
      - source_id: UniProtKB:Q2M2I8
        source_label: human AAK1 target record
        source_status: CIRCULAR_OR_REDUNDANT
        comment: >-
          The target protein is listed as its own IBA source and therefore cannot
          serve as independent evidence for propagation.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: Thus, these results demonstrate that AAK1 interacts directly with α-adaptin in vitro.
- term:
    id: GO:2000369
    label: regulation of clathrin-dependent endocytosis
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: involved_in
  supporting_entities:
  - PANTHER:PTN000539992
  - RGD:1305520
  - UniProtKB:F1SPM8
  - UniProtKB:Q2M2I8
  review:
    summary: >-
      Phylogenetic assignment of regulation of clathrin-dependent endocytosis.
      This is the central biological role of AAK1 and is supported by multiple
      direct perturbation and biochemical studies.
    action: ACCEPT
    reason: >-
      AAK1 regulates cargo capture and coated-pit maturation through AP2M1
      phosphorylation and has experimentally demonstrated effects on transferrin,
      LRP, and other cargo internalization (PMID:11877457, PMID:11877461,
      PMID:12952931, PMID:17494869, PMID:30605688, PMID:34315807). The
      PMID:12952931 uptake phenotype came from AAK1 overexpression, while about 80%
      knockdown produced no detectable uptake phenotype, so it is corroborating but
      not decisive evidence. The human target appears among the IBA sources and is
      treated as redundant rather than independent evidence.
    additional_reference_ids:
    - PMID:11877457
    - PMID:11877461
    - PMID:12952931
    - PMID:17494869
    - PMID:30605688
    - PMID:34315807
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: PANTHER:PTN000539992
        source_label: PANTHER AAK1 family node
        source_status: SOURCE_WEAK_OR_INFERRED
        comment: The family node is the correlated inference carrier.
      - source_id: RGD:1305520
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: Rat Aak1 supports conservation of the endocytic regulatory role.
      - source_id: UniProtKB:F1SPM8
        source_label: pig AAK1
        source_status: SUPPORTS_TRANSFER
        comment: Pig AAK1 is an orthologous donor supporting the transfer.
      - source_id: UniProtKB:Q2M2I8
        source_label: human AAK1 target record
        source_status: CIRCULAR_OR_REDUNDANT
        comment: >-
          Self-support in the IBA source set is redundant and is not counted as an
          independent line of evidence.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        Together, these results provide strong evidence that AAK1 is the endogenous mu 2 kinase and plays a regulatory role in clathrin-mediated endocytosis.
- term:
    id: GO:0004672
    label: protein kinase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: enables
  supporting_entities:
  - InterPro:IPR008271
  review:
    summary: >-
      InterPro electronically maps the AAK1 kinase-domain signature to the broad
      term protein kinase activity. The catalytic class is correct, but a more
      informative serine/threonine-specific term is already supported directly.
    action: MODIFY
    reason: >-
      AAK1 is not merely an unspecified protein kinase; biochemical and structural
      evidence establishes serine/threonine protein kinase activity. Replace the
      broad parent with GO:0004674, consolidating with existing GO:0004674 rows.
    proposed_replacement_terms:
    - id: GO:0004674
      label: protein serine/threonine kinase activity
    additional_reference_ids:
    - PMID:11877457
    - PMID:11877461
    - PMID:26853940
    propagation_review:
      root_cause: TERM_SCOPING_PROBLEM
      failure_modes:
      - GRANULARITY_MISMATCH
      source_entities:
      - source_id: InterPro:IPR008271
        source_label: protein kinase active-site signature
        source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET
        comment: >-
          The signature supports kinase activity but the electronic mapping is
          broader than the experimentally established serine/threonine specificity;
          it is also correlated with the other kinase-domain mappings.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        Here we identify and characterize a novel member of the Prk/Ark family of serine/threonine kinases, adaptor-associated kinase (AAK)1.
- term:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000003
  qualifier: enables
  supporting_entities:
  - EC:2.7.11.1
  review:
    summary: >-
      EC-to-GO electronic assignment of protein serine/threonine kinase activity.
      It matches AAK1's experimentally established catalytic class.
    action: ACCEPT
    reason: >-
      EC 2.7.11.1 is correctly mapped to the core molecular function, which is
      independently supported by direct AP2M1 phosphorylation and human AAK1 kinase
      structures. The EC mapping is corroborative and correlated with the other
      electronic kinase-domain rows, not a separate experiment.
    additional_reference_ids:
    - PMID:11877457
    - PMID:26853940
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: EC:2.7.11.1
        source_label: non-specific serine/threonine protein kinase EC class
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The EC mapping has the correct catalytic scope; it is an electronic
          classification correlated with the InterPro and Rhea mappings.
- term:
    id: GO:0005524
    label: ATP binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: enables
  supporting_entities:
  - InterPro:IPR000719
  review:
    summary: >-
      InterPro kinase-domain mapping to ATP binding. ATP binding is mechanistically
      correct for AAK1 but is generic relative to its informative catalytic activity.
    action: KEEP_AS_NON_CORE
    reason: >-
      The AAK1 kinase domain contains the conserved ATP-binding pocket and direct
      structural work resolves inhibitor binding in that pocket (PMID:26853940).
      Retain the annotation as a valid cofactor-binding property, but do not elevate
      generic ATP binding to a core functional summary.
    additional_reference_ids:
    - PMID:26853940
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: InterPro:IPR000719
        source_label: protein kinase domain
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The domain supports ATP binding, but this electronic row is correlated
          with the other kinase-domain mappings and adds no independent experiment.
    supported_by:
    - reference_id: PMID:26853940
      supporting_text: >-
        Here we report the first high-resolution structures of kinases AAK1 and BIKE in complex with two drug candidates.
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  supporting_entities:
  - UniProtKB-SubCell:SL-0039
  review:
    summary: >-
      Electronic localization to the plasma membrane derived from the UniProt
      subcellular-location vocabulary. This is consistent with AAK1 action at
      plasma-membrane clathrin-coated pits and with direct human-cell imaging.
    action: ACCEPT
    reason: >-
      AAK1 acts peripherally at the plasma membrane during cargo internalization.
      Direct RPE-cell imaging found broad plasma-membrane AAK1 distribution
      (PMID:42082516), independently supporting the location. The vocabulary mapping
      is generated from the same UniProt target record and is therefore correlated,
      not an independent localization experiment.
    additional_reference_ids:
    - PMID:42082516
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB-SubCell:SL-0039
        source_label: UniProt cell membrane vocabulary term
        source_status: CIRCULAR_OR_REDUNDANT
        comment: >-
          This IEA maps the target record's own subcellular-location statement; it
          is valid but self-derived and correlated with other UniProt location rows.
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
- term:
    id: GO:0005905
    label: clathrin-coated pit
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  supporting_entities:
  - UniProtKB-SubCell:SL-0069
  review:
    summary: >-
      Electronic localization to clathrin-coated pits from the UniProt
      subcellular-location vocabulary. This is a principal site at which AAK1
      controls AP2 and cargo capture.
    action: ACCEPT
    reason: >-
      Direct localization and biochemical work place AAK1 with AP2 and clathrin at
      coated pits (PMID:11877461), while clathrin activates AAK1
      (PMID:14617351). The mapping is correct but derives from the same UniProt
      record, so it should not be counted as independent evidence.
    additional_reference_ids:
    - PMID:11877461
    - PMID:14617351
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB-SubCell:SL-0069
        source_label: UniProt clathrin-coated pit vocabulary term
        source_status: CIRCULAR_OR_REDUNDANT
        comment: >-
          This is a self-derived vocabulary mapping from the AAK1 UniProt record,
          correlated with its other mapped localization rows.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        AAK1 also colocalized to endocytic structures in nonneuronal cells.
- term:
    id: GO:0098793
    label: presynapse
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  supporting_entities:
  - UniProtKB-SubCell:SL-0516
  review:
    summary: >-
      Electronic presynapse localization mapped from the UniProt record. The
      location is plausible from neuronal ortholog evidence but is tissue-specific.
    action: KEEP_AS_NON_CORE
    reason: >-
      Rat-neuron experiments reported enrichment at presynaptic terminals
      (PMID:11877461). Retain this as contextual neuronal localization rather than
      a general core location of human AAK1. Because the IEA is mapped from the
      target record's own location statement, it is correlated/self-derived.
    additional_reference_ids:
    - PMID:11877461
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: UniProtKB-SubCell:SL-0516
        source_label: UniProt presynapse vocabulary term
        source_status: CIRCULAR_OR_REDUNDANT
        comment: >-
          The source is the target record's own vocabulary assertion and is not an
          independent human presynaptic localization experiment.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: In neuronal cells, AAK1 is enriched at presynaptic terminals
- term:
    id: GO:0106310
    label: protein serine kinase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000116
  qualifier: enables
  supporting_entities:
  - RHEA:17989
  review:
    summary: >-
      Rhea electronically maps the serine-specific protein phosphorylation reaction
      to the current protein serine kinase activity term. This is one valid
      subactivity within AAK1's broader serine/threonine kinase activity.
    action: ACCEPT
    reason: >-
      RHEA:17989 directly supports the serine-specific electronic mapping, and this
      annotation can coexist with the broader GO:0004674 rows. AAK1 also has strong
      threonine-substrate evidence, but current GO has no separate primary protein
      threonine kinase term: GO:0106311 is an alternate ID of GO:0004674, not a
      usable sibling term. Retain the valid serine subactivity rather than
      generalizing it or writing an alternate identifier.
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: RHEA:17989
        source_label: L-seryl-protein phosphorylation reaction
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The reaction directly supports the serine subactivity. It is correlated
          with the UniProt catalytic record and does not negate AAK1's independently
          supported threonine activity.
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:22645275
  qualifier: enables
  supporting_entities:
  - UniProtKB:Q9NQ11
  review:
    summary: >-
      GOA/IntAct records AAK1 as an ATP13A2 interactor from a yeast two-hybrid
      screen. The cached abstract describes the 43-interactor screen but does not
      name AAK1, and the full text is unavailable locally, so the specific pair
      cannot be independently verified here.
    action: MARK_AS_OVER_ANNOTATED
    reason: >-
      Retain and defer to the experimental interaction provenance; this action does
      not reject or question the AAK1-ATP13A2 pair. It concerns only GO:0005515,
      which is an uninformative generic molecular-function term and does not capture
      the interaction partner, mechanism, or biological role. Mark the term as
      over-annotated while recording that the pair-level evidence remains
      unverified from the locally accessible abstract.
    supported_by:
    - reference_id: PMID:22645275
      supporting_text: >-
        Utilizing a split-ubiquitin membrane yeast two-hybrid system that was developed to identify interacting partners of full-length integral membrane proteins, we identified 43 novel interactors
- term:
    id: GO:0061024
    label: membrane organization
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-199991
  qualifier: involved_in
  review:
    summary: >-
      Reactome assigns AAK1 to the broad membrane-trafficking umbrella and maps that
      participation to membrane organization. This captures pathway context but is
      broader than AAK1's specific clathrin-mediated endocytic role.
    action: KEEP_AS_NON_CORE
    reason: >-
      Membrane organization is defensible as a high-level consequence of AAK1's
      endocytic function, but the core role is regulation of clathrin-dependent
      endocytosis through AP2M1 phosphorylation. Retain this TAS annotation as broad
      pathway context rather than as a defining function.
- term:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8856813
  qualifier: enables
  review:
    summary: >-
      Reactome represents AAK1 phosphorylating the AP-2 mu subunit and maps the event
      to protein serine/threonine kinase activity. This matches the core catalytic
      function established experimentally.
    action: ACCEPT
    reason: >-
      The pathway event is consistent with direct biochemical evidence that AAK1
      phosphorylates AP2M1 at Thr-156 (PMID:11877457). Although the TAS is model
      based, its catalytic assertion is correctly scoped and independently supported.
    additional_reference_ids:
    - PMID:11877457
    supported_by:
    - reference_id: PMID:11877457
      supporting_text: >-
        Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IDA
  original_reference_id: GO_REF:0000052
  qualifier: located_in
  review:
    summary: >-
      Human Protein Atlas immunofluorescence places AAK1 at the plasma membrane.
      This agrees with its role at plasma-membrane clathrin-coated pits.
    action: ACCEPT
    reason: >-
      The direct human-cell localization is biologically consistent with AAK1 cargo
      internalization and with independent live-cell evidence of plasma-membrane
      distribution (PMID:42082516). Retain the experimentally assigned location.
    additional_reference_ids:
    - PMID:42082516
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:F1MH24
  review:
    summary: >-
      Plasma-membrane localization transferred by curator-reviewed similarity from
      bovine AAK1. This is conserved and independently supported in human cells.
    action: ACCEPT
    reason: >-
      The bovine ortholog is an appropriate donor and direct imaging of human AAK1
      supports plasma-membrane distribution (PMID:42082516). The same donor is reused
      for multiple location rows, so those ISS annotations are correlated rather than
      independent observations.
    additional_reference_ids:
    - PMID:42082516
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB:F1MH24
        source_label: bovine AAK1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The ortholog supports transfer; reuse of this single donor for plasma
          membrane and coated pit makes the inferred rows correlated.
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        In contrast, AAK1 is broadly distributed across the PM, with 40% lower fluorescence intensity in CCPs in comparison to BMP2K.
- term:
    id: GO:0098793
    label: presynapse
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:P0C1X8
  review:
    summary: >-
      Presynapse localization transferred from rat Aak1. The inference is plausible
      and supported in rat neurons, but it is a neuronal-context location.
    action: KEEP_AS_NON_CORE
    reason: >-
      Preserve the ortholog transfer because rat-neuron experiments place AAK1 at
      presynaptic terminals (PMID:11877461), while recognizing that the human row is
      model-derived and should not define AAK1's general cellular location. This and
      other P0C1X8 transfers reuse one donor and are correlated.
    additional_reference_ids:
    - PMID:11877461
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: UniProtKB:P0C1X8
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          Rat Aak1 supports the neuronal localization; reuse of this donor across
          kinase and neuronal-location rows does not constitute independent evidence.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: In neuronal cells, AAK1 is enriched at presynaptic terminals
- term:
    id: GO:0005905
    label: clathrin-coated pit
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:F1MH24
  review:
    summary: >-
      Clathrin-coated-pit localization transferred from bovine AAK1. The location is
      central to AAK1's endocytic role and is independently observed in human cells.
    action: ACCEPT
    reason: >-
      AAK1 colocalizes with AP2 and clathrin at coated-pit structures and is activated
      by assembled clathrin (PMID:11877461, PMID:14617351). The bovine donor is sound,
      but its reuse with the plasma-membrane ISS row makes the evidence correlated.
    additional_reference_ids:
    - PMID:11877461
    - PMID:14617351
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB:F1MH24
        source_label: bovine AAK1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          Bovine AAK1 is a suitable orthologous donor; this is the same donor used
          for the plasma-membrane transfer and is not an independent second source.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        AAK1 also colocalized to endocytic structures in nonneuronal cells.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8856808
  qualifier: located_in
  review:
    summary: >-
      The Reactome AP-2/clathrin recruitment event models AAK1 as cytosolic before
      or during recruitment to the endocytic machinery.
    action: KEEP_AS_NON_CORE
    reason: >-
      A cytosolic pool is plausible for this peripheral-membrane kinase, but this
      TAS is generated from a shared Reactome clathrin-mediated-endocytosis model.
      The 17 Reactome cytosol rows are correlated pathway-context assertions, not
      independent localization experiments; retain each as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8856813
  qualifier: located_in
  review:
    summary: >-
      The Reactome AP2M1 phosphorylation event assigns its AAK1 participant to the
      cytosol.
    action: KEEP_AS_NON_CORE
    reason: >-
      Cytosolic availability is compatible with recruitment of AAK1 to coated pits,
      but the location is model context rather than a direct assay. It is correlated
      with the other 16 cytosol tuples emitted from the same Reactome pathway and is
      retained as non-core.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866283
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled recruitment of GPCRs to
      clathrin-coated pits.
    action: KEEP_AS_NON_CORE
    reason: >-
      This is a correlated participant-location assertion from the shared Reactome
      endocytosis model, not an independent localization experiment or a distinct
      AAK1 function. Retain it as non-core pathway context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8867754
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the modeled F-/N-BAR protein recruitment
      phase of coated-pit maturation.
    action: KEEP_AS_NON_CORE
    reason: >-
      The cytosolic location is biologically compatible with a recruitable peripheral
      kinase, but this tuple is one of 17 correlated annotations generated from the
      same pathway model. Keep it as non-core context rather than independent support.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8867756
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in a modeled nascent coated-pit cargo/CLASP
      recruitment event.
    action: KEEP_AS_NON_CORE
    reason: >-
      This pathway-derived location is plausible but does not establish a separate
      localization result. It is correlated with the other Reactome cytosol rows and
      is retained as non-core endocytic context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868071
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the clathrin-dependent PIK3C2A
      recruitment event.
    action: KEEP_AS_NON_CORE
    reason: >-
      The TAS reflects a shared pathway compartment rather than an AAK1-specific
      localization experiment. It is correlated with the 16 other Reactome cytosol
      rows and is retained only as non-core pathway context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868072
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled PIK3C2A phosphoinositide
      phosphorylation stage of coated-pit maturation.
    action: KEEP_AS_NON_CORE
    reason: >-
      This is a pathway-participant compartment assignment, not direct evidence that
      independently localizes AAK1. Treat it as correlated with the other Reactome
      cytosol tuples and retain it as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868230
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled SNX9/actin-machinery recruitment
      stage of clathrin-mediated endocytosis.
    action: KEEP_AS_NON_CORE
    reason: >-
      The location derives from the common pathway model and is not an independent
      AAK1 localization assay. It is correlated with the other 16 cytosol rows and
      should remain non-core.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868236
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in a modeled BAR-protein/dynamin recruitment
      event.
    action: KEEP_AS_NON_CORE
    reason: >-
      Cytosol is plausible for the unbound pool, but this TAS is a correlated
      pathway-context assertion rather than direct localization evidence. Retain it
      as non-core.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868648
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled synaptojanin lipid-hydrolysis
      stage of coated-vesicle formation.
    action: KEEP_AS_NON_CORE
    reason: >-
      This tuple inherits the pathway compartment and is not independent evidence
      for AAK1 localization or synaptojanin regulation. It is correlated with the
      other Reactome cytosol rows and is retained as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868651
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the modeled endophilin/synaptojanin
      recruitment event.
    action: KEEP_AS_NON_CORE
    reason: >-
      The assignment is a shared pathway-location statement, not an AAK1-specific
      experiment. It is correlated with the other 16 Reactome cytosol annotations
      and is kept only as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868658
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled HSPA8-driven vesicle-uncoating
      event.
    action: KEEP_AS_NON_CORE
    reason: >-
      This is correlated compartment context from a broader endocytosis model, not
      evidence that AAK1 directly performs or controls HSPA8 ATP hydrolysis. Retain
      the cytosol tuple as non-core.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868659
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the modeled recruitment of auxilins to
      clathrin-coated vesicles.
    action: KEEP_AS_NON_CORE
    reason: >-
      The TAS derives from the shared pathway compartment rather than a direct AAK1
      localization assay. It is correlated with the other Reactome cytosol tuples
      and is retained as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868660
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled auxilin/HSPA8 recruitment event.
    action: KEEP_AS_NON_CORE
    reason: >-
      This participant-location row is correlated with the common Reactome pathway
      model and does not provide an independent localization observation. Retain it
      as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8868661
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the modeled dynamin-mediated vesicle
      scission event.
    action: KEEP_AS_NON_CORE
    reason: >-
      The tuple inherits a pathway compartment and should not be interpreted as
      separate evidence that AAK1 controls dynamin catalysis. It is correlated with
      the other 16 cytosol annotations and is retained as non-core context.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8871193
  qualifier: located_in
  review:
    summary: >-
      Reactome assigns cytosolic AAK1 in the modeled dissociation of AAK1 and
      dephosphorylation of AP2M1.
    action: KEEP_AS_NON_CORE
    reason: >-
      Cytosolic release after coated-pit action is mechanistically plausible, but
      this is model-derived and correlated with the other Reactome cytosol rows. It
      is retained as non-core rather than treated as direct localization evidence.
- term:
    id: GO:0005829
    label: cytosol
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8871194
  qualifier: located_in
  review:
    summary: >-
      Reactome places AAK1 in the cytosol in the modeled RAB5/GAPVD1 binding phase
      of the AP-2 endocytic pathway.
    action: KEEP_AS_NON_CORE
    reason: >-
      The TAS is a correlated pathway-participant compartment assignment, not a
      distinct AAK1 localization experiment. Retain it as non-core context with the
      other Reactome cytosol tuples.
- term:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: enables
  supporting_entities:
  - UniProtKB:P0C1X8
  review:
    summary: >-
      Protein serine/threonine kinase activity transferred from rat Aak1 by
      curator-reviewed similarity. This is the defining catalytic activity and is
      independently established for human AAK1.
    action: ACCEPT
    reason: >-
      The rat ortholog is an appropriate donor, and direct biochemical and structural
      studies of human AAK1 confirm an active serine/threonine kinase
      (PMID:11877457, PMID:11877461, PMID:26853940). The same donor is reused for
      several localization rows, so those transfers are correlated rather than
      independent support.
    additional_reference_ids:
    - PMID:11877457
    - PMID:11877461
    - PMID:26853940
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB:P0C1X8
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          Rat Aak1 is a suitable orthologous donor; reuse of this one donor across
          kinase, presynapse, leading-edge, and bouton rows is correlated evidence.
    supported_by:
    - reference_id: PMID:11877457
      supporting_text: >-
        Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
- term:
    id: GO:0005112
    label: Notch binding
  evidence_type: IDA
  original_reference_id: PMID:21464124
  qualifier: enables
  review:
    summary: >-
      AAK1 directly binds the membrane-tethered activated form of NOTCH1, but not the
      inactive full-length receptor, in a ligand-activation/endocytic context.
    action: KEEP_AS_NON_CORE
    reason: >-
      The IDA is directly supported and more informative than generic protein
      binding. Retain it as a context-specific interaction that helps route activated
      NOTCH1, rather than treating Notch binding as the defining molecular function
      of AAK1.
    supported_by:
    - reference_id: PMID:21464124
      supporting_text: >-
        We show here that AAK1, the adaptor-associated kinase 1, directly interacts with the membrane-tethered active form of Notch released by metalloprotease cleavage.
- term:
    id: GO:0006468
    label: protein phosphorylation
  evidence_type: IDA
  original_reference_id: PMID:17494869
  qualifier: involved_in
  review:
    summary: >-
      AAK1L and AAK1S were tested in kinase assays against the AP-2 mu2 subunit,
      whose established AAK1 phosphosite is Thr-156.
    action: MODIFY
    reason: >-
      The study directly assayed both human splice forms and found basal and
      clathrin-stimulated AP2M1 kinase activity. Replace the broad process term with
      GO:0018107 peptidyl-threonine phosphorylation, supported by direct mapping of
      the AAK1-dependent AP2M1 site to Thr-156 in PMID:11877457.
    proposed_replacement_terms:
    - id: GO:0018107
      label: peptidyl-threonine phosphorylation
    additional_reference_ids:
    - PMID:11877457
    supported_by:
    - reference_id: PMID:17494869
      supporting_text: >-
        However, in vitro kinase assays reveal little difference between AAK1 isoforms in their basal or clathrin-stimulated kinase activity toward the AP-2 micro2 subunit.
    - reference_id: PMID:11877457
      supporting_text: >-
        Phosphorylation of mu 2 occurs at a single threonine residue (Thr-156) and is mediated by the newly discovered adaptor-associated kinase, AAK1, which copurifies with AP2.
- term:
    id: GO:0006468
    label: protein phosphorylation
  evidence_type: IDA
  original_reference_id: PMID:18657069
  qualifier: involved_in
  review:
    summary: >-
      The source reports AAK1-dependent phosphorylation of NUMB and identifies T102
      as the relevant site. A 2026 biochemical study did not detect phosphorylation
      of a NUMB T102 peptide, creating substrate-site-specific counterevidence.
    action: MODIFY
    reason: >-
      Replace the broad process term with GO:0018107 peptidyl-threonine
      phosphorylation. The original study reports a threonine phosphosite, while
      AP2M1 T156 and newer PDLIM5/Talin1 sites independently establish AAK1-mediated
      threonine phosphorylation. This replacement does not endorse NUMB T102 as a
      physiological substrate: that specific assignment is disputed by
      PMID:42082516.
    proposed_replacement_terms:
    - id: GO:0018107
      label: peptidyl-threonine phosphorylation
    additional_reference_ids:
    - PMID:11877457
    - PMID:42082516
    supported_by:
    - reference_id: PMID:18657069
      supporting_text: >-
        In this study, we show that Numb binds to and is phosphorylated by adaptor-associated kinase 1 (AAK1), a key endocytic kinase.
    - reference_id: PMID:42082516
      supporting_text: >-
        This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
- term:
    id: GO:0030136
    label: clathrin-coated vesicle
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:F1SPM8
  review:
    summary: >-
      Clathrin-coated-vesicle localization transferred from pig AAK1. This is
      consistent with AAK1 association with AP2, clathrin, and isolated coated
      vesicles during its core endocytic function.
    action: ACCEPT
    reason: >-
      The pig ortholog is an appropriate donor, and biochemical fractionation of
      mammalian tissues independently recovered AAK1 with clathrin-coated vesicles
      (PMID:11877461). This donor also supports an IBA process row, making those
      propagated assertions correlated rather than independent.
    additional_reference_ids:
    - PMID:11877461
    propagation_review:
      root_cause: NO_FAILURE_CORE
      source_entities:
      - source_id: UniProtKB:F1SPM8
        source_label: pig AAK1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          Pig AAK1 supports the conserved coated-vesicle location; reuse of this
          donor in the IBA endocytosis row is correlated evidence.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        AAK1 is found associated with isolated APs (lane 1), clathrin (lane 2), and CCVs (lane 3), and CCVs from rat liver (lane 4).
- term:
    id: GO:0031252
    label: cell leading edge
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:P0C1X8
  review:
    summary: >-
      Cell-leading-edge localization transferred from rat Aak1. Direct human-cell
      work supports leading-edge/endocytic and focal-adhesion recruitment during
      migration, but this is context-dependent.
    action: KEEP_AS_NON_CORE
    reason: >-
      Human HeLa imaging reported AAK1 enrichment at the leading edge
      (PMID:11877461), and newer RPE-cell work links AAK1 recruitment to focal
      adhesion turnover and migration (PMID:42082516). Retain this localization as a
      non-core migratory-cell context. The rat donor is reused across several ISS
      rows and thus does not create independent evidence for each.
    additional_reference_ids:
    - PMID:11877461
    - PMID:42082516
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: UniProtKB:P0C1X8
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          Rat Aak1 is an appropriate donor, but this single donor is shared with the
          kinase, presynapse, and terminal-bouton ISS rows.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: >-
        Interestingly, in migrating cells AAK1 was highly enriched at the leading edge
- term:
    id: GO:0032880
    label: regulation of protein localization
  evidence_type: IDA
  original_reference_id: PMID:18657069
  qualifier: involved_in
  review:
    summary: >-
      AAK1 perturbation changes NUMB distribution between the plasma membrane and
      perinuclear endosomes, directly supporting regulation of protein localization.
    action: KEEP_AS_NON_CORE
    reason: >-
      Preserve the reported localization phenotype even though the proposed direct
      phosphorylation of NUMB T102 is challenged by PMID:42082516. The redistribution
      result supports a real, substrate/context-specific regulatory output, but it is
      secondary to AAK1's core AP2/endocytic function.
    additional_reference_ids:
    - PMID:42082516
    supported_by:
    - reference_id: PMID:18657069
      supporting_text: >-
        We find that AAK1 redistributes Numb to perinuclear endosomes when overexpressed, while kinase depletion causes Numb to accumulate at the plasma membrane.
    - reference_id: PMID:42082516
      supporting_text: >-
        This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
- term:
    id: GO:0035612
    label: AP-2 adaptor complex binding
  evidence_type: IDA
  original_reference_id: PMID:17494869
  qualifier: enables
  review:
    summary: >-
      The source supports association of AAK1 isoforms with the AP-2 machinery and
      assays AP2M1 as their kinase substrate.
    action: ACCEPT
    reason: >-
      Although the locally cached source exposes only the abstract and does not spell
      out every binding assay, the experimental curator had access to the full study.
      Direct AP-2/alpha-adaptin binding was independently demonstrated in
      PMID:11877461, so the informative AP-2-complex-binding term is retained.
    additional_reference_ids:
    - PMID:11877461
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: Thus, these results demonstrate that AAK1 interacts directly with α-adaptin in vitro.
- term:
    id: GO:0043195
    label: terminal bouton
  evidence_type: ISS
  original_reference_id: GO_REF:0000024
  qualifier: located_in
  supporting_entities:
  - UniProtKB:P0C1X8
  review:
    summary: >-
      Terminal-bouton localization transferred from rat Aak1. It is compatible with
      reported presynaptic-terminal enrichment but is a neuronal model-derived
      sublocation rather than a general human AAK1 location.
    action: KEEP_AS_NON_CORE
    reason: >-
      Keep the ortholog transfer as contextual neuronal localization. The cached
      experimental evidence supports presynaptic-terminal enrichment
      (PMID:11877461) but not a uniquely required terminal-bouton role, and the same
      rat donor is reused across multiple ISS rows.
    additional_reference_ids:
    - PMID:11877461
    propagation_review:
      root_cause: NO_FAILURE_NON_CORE
      source_entities:
      - source_id: UniProtKB:P0C1X8
        source_label: rat Aak1
        source_status: SUPPORTS_TRANSFER
        comment: >-
          The donor supports neuronal terminal localization, but this is the same
          source reused for presynapse, kinase, and leading-edge transfers.
    supported_by:
    - reference_id: PMID:11877461
      supporting_text: In neuronal cells, AAK1 is enriched at presynaptic terminals
- term:
    id: GO:0045747
    label: positive regulation of Notch signaling pathway
  evidence_type: IDA
  original_reference_id: PMID:21464124
  qualifier: involved_in
  review:
    summary: >-
      AAK1 promotes activated NOTCH1 routing and transcriptional output upstream of
      gamma-secretase, supporting positive regulation of Notch signaling in the
      tested ligand-activation context.
    action: KEEP_AS_NON_CORE
    reason: >-
      The direct perturbation evidence supports a positive effect on mammalian Notch
      signaling. Retain it as a context-specific signaling consequence of AAK1's
      endocytic/adaptor activity, rather than as the kinase's universal core process.
    supported_by:
    - reference_id: PMID:21464124
      supporting_text: >-
        Moreover, shRNA-mediated AAK1 depletion decreases Notch transcriptional activity in coculture experiments.
- term:
    id: GO:0050821
    label: protein stabilization
  evidence_type: IDA
  original_reference_id: PMID:21464124
  qualifier: involved_in
  review:
    summary: >-
      AAK1 stabilizes the membrane-tethered activated NOTCH1 fragment and its
      monoubiquitinated counterpart upstream of gamma-secretase cleavage.
    action: KEEP_AS_NON_CORE
    reason: >-
      The IDA is supported for the specific activated-NOTCH1 experimental context.
      Protein stabilization is a secondary, substrate-specific output of AAK1
      adaptor/endocytic activity and should not be treated as its defining process.
    supported_by:
    - reference_id: PMID:21464124
      supporting_text: >-
        Active AAK1 acts upstream of the γ-secretase cleavage by stabilizing both the membrane-tethered activated form of Notch and its monoubiquitinated counterpart.
- term:
    id: GO:2000369
    label: regulation of clathrin-dependent endocytosis
  evidence_type: IMP
  original_reference_id: PMID:17494869
  qualifier: involved_in
  review:
    summary: >-
      AAK1L-domain overexpression and AAK1 depletion perturb transferrin
      internalization and recycling, directly demonstrating regulation at multiple
      steps of the clathrin-dependent endocytic pathway.
    action: ACCEPT
    reason: >-
      This IMP annotation captures the central biological role established by
      perturbation of AAK1 constructs and abundance. Cargo and direction of effect
      vary with experimental context, so the sign-neutral regulatory GO term is
      appropriately scoped.
    supported_by:
    - reference_id: PMID:17494869
      supporting_text: >-
        These observations suggest that AAK1 functions at multiple steps of the endosomal pathway by regulating transferrin internalization and its rapid recycling back to the plasma membrane from early/sorting endosome.
- term:
    id: GO:2000369
    label: regulation of clathrin-dependent endocytosis
  evidence_type: IDA
  original_reference_id: PMID:18657069
  qualifier: involved_in
  review:
    summary: >-
      AAK1 perturbation and NUMB localization/mutant experiments support regulation
      of cargo-selective clathrin-dependent endocytosis and coated-pit maturation.
    action: ACCEPT
    reason: >-
      Preserve the broad endocytic-regulation annotation: the reported NUMB
      redistribution and transferrin/LDL uptake phenotypes support it, and AAK1's
      endocytic role is independently established (PMID:11877461, PMID:17494869).
      PMID:42082516 disputes direct phosphorylation of the proposed NUMB T102 peptide,
      so that specific mechanism is treated cautiously without discarding the
      phenotype or the broader process.
    additional_reference_ids:
    - PMID:11877461
    - PMID:17494869
    - PMID:42082516
    supported_by:
    - reference_id: PMID:18657069
      supporting_text: >-
        Overexpression of a Numb point mutant (T102A) that lacks the AAK1 phosphorylation site potently disrupts transferrin and low-density lipoprotein internalization but does not impact EGF uptake.
    - reference_id: PMID:42082516
      supporting_text: >-
        This refined consensus sequence also explains why previously proposed substrates Dab2 T109, Arh T104, NUMB T102, and AP1μ T14426 showed no detectable phosphorylation in our in-vitro experiments
- term:
    id: GO:0030276
    label: clathrin binding
  evidence_type: IDA
  original_reference_id: PMID:17494869
  qualifier: enables
  isoform: Q2M2I8-1
  review:
    summary: >-
      NEW annotation. The long AAK1 isoform contains a second C-terminal
      clathrin-binding region whose direct interaction with clathrin was tested in
      protein-interaction assays.
    action: NEW
    reason: >-
      Clathrin binding is an informative molecular function that helps recruit and
      stimulate AAK1 in the coated-pit machinery. The isoform field records that the
      tested CBD2 construct came from long isoform AAK1L/Q2M2I8-1; it does not claim
      that clathrin binding is unique to this isoform.
    supported_by:
    - reference_id: PMID:17494869
      supporting_text: >-
        Protein interaction studies demonstrate that AAK1L CBD2 directly binds clathrin.
- term:
    id: GO:2001137
    label: positive regulation of endocytic recycling
  evidence_type: IMP
  original_reference_id: PMID:17494869
  qualifier: involved_in
  isoform: Q2M2I8-1
  review:
    summary: >-
      NEW annotation. AAK1 depletion or interference with its second
      clathrin-binding region impairs transferrin recycling from early/sorting
      endosomes, and independent depletion experiments delay short-loop beta3
      integrin recycling to the cell surface.
    action: NEW
    reason: >-
      Loss-of-function evidence for two cargos supports a positive contribution to
      endocytic recycling, a process not captured by the existing annotation to
      regulation of clathrin-dependent endocytosis. The isoform field records the
      tested long AAK1L/Q2M2I8-1 constructs without claiming that the shorter isoform
      cannot contribute.
    additional_reference_ids:
    - PMID:23781025
    supported_by:
    - reference_id: PMID:17494869
      supporting_text: >-
        Surprisingly, CBD2 overexpression or AAK1 depletion by RNA interference significantly impairs transferrin recycling from the early/sorting endosome.
    - reference_id: PMID:23781025
      supporting_text: >-
        siRNA-mediated depletion of either factor delays short-loop β3 integrin recycling from the early endosome back to the cell surface.
- term:
    id: GO:0005925
    label: focal adhesion
  evidence_type: IDA
  original_reference_id: PMID:42082516
  qualifier: located_in
  review:
    summary: >-
      NEW annotation. Live-cell imaging places AAK1 at mature, tension-bearing
      focal adhesions in collagen-adherent human RPE cells.
    action: NEW
    reason: >-
      The ECM-dependent localization was supported by paxillin colocalization and
      multiple illumination, membrane, and attachment controls. This is the site of
      AAK1's directly demonstrated adhesion-turnover activity.
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        Thus, AAK1 is specifically recruited to mature, tension-bearing FAs and thereby positioned to efficiently phosphorylate substrates PDLIM5 and Talin1.
- term:
    id: GO:0120183
    label: positive regulation of focal adhesion disassembly
  evidence_type: IMP
  original_reference_id: PMID:42082516
  qualifier: involved_in
  review:
    summary: >-
      NEW annotation. AAK1 deletion increases focal-adhesion density and lifetime
      and slows paxillin exchange, while kinase- and phosphosite-dependent analyses
      support accelerated release of focal-adhesion components.
    action: NEW
    reason: >-
      Human-cell loss-of-function, live imaging, rescue, and phosphomutant evidence
      directly supports a positive regulatory role in focal-adhesion disassembly.
      This is a core AAK1 function distinct from its cargo-trafficking role.
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        Because rapid paxillin turnover is essential for efficient cell migration, these data position AAK1 as a positive regulator of FA disassembly and motility.
- term:
    id: GO:0030335
    label: positive regulation of cell migration
  evidence_type: IMP
  original_reference_id: PMID:42082516
  qualifier: involved_in
  review:
    summary: >-
      NEW annotation. AAK1 knockout or knockdown slows human RPE-cell migration,
      with corroborating knockdown phenotypes in SH-SY5Y and MDA-MB-231 cells.
    action: NEW
    reason: >-
      Genetic loss and kinase-inhibitor phenotypes establish a positive,
      kinase-dependent contribution to cell migration. BMP2K loss did not reproduce
      the RPE phenotype, supporting an AAK1-specific core role.
    supported_by:
    - reference_id: PMID:42082516
      supporting_text: >-
        AAK1 deletion or siRNA-mediated knockdown reproducibly impaired RPE migration by ~25% (Fig. 3d, e and Supplementary Fig. 1h), whereas BMP2K loss had no measurable effect, revealing a specific and non-redundant role for AAK1 in cell motility.
- term:
    id: GO:0090090
    label: negative regulation of canonical Wnt signaling pathway
  evidence_type: IMP
  original_reference_id: PMID:30605688
  qualifier: involved_in
  review:
    summary: >-
      NEW annotation. AAK1 loss or inhibition increases canonical WNT output in
      multiple human cell models, while AAK1 promotes LRP6 endocytosis and
      lysosomal degradation.
    action: NEW
    reason: >-
      Perturbation and mechanistic evidence support negative regulation of canonical
      WNT signaling. This is retained as a context- and cargo-specific output of the
      endocytic machinery, not as a universal core function of AAK1.
    supported_by:
    - reference_id: PMID:30605688
      supporting_text: >-
        Together, these results suggest that AAK1 inhibits WNT signaling by inducing CME endocytosis and lysosomal degradation of LRP6.
- term:
    id: GO:0160020
    label: positive regulation of ferroptosis
  evidence_type: IMP
  original_reference_id: PMID:41407700
  qualifier: involved_in
  review:
    summary: >-
      NEW annotation. In induced ferroptosis models, AAK1 loss reduces lipid
      peroxidation and ferroptotic death, whereas activated AAK1 drives
      AP2M1-dependent TFR1 internalization and iron accumulation.
    action: NEW
    reason: >-
      Genetic and mechanistic evidence supports positive regulation of ferroptosis
      in the tested cancer-cell and xenograft settings. This stimulus-dependent,
      cargo-specific role is non-core rather than a constitutive output of AAK1.
    supported_by:
    - reference_id: PMID:41407700
      supporting_text: >-
        Mechanistically, activated AAK1 phosphorylates AP2M1, which facilitates the recruitment of clathrin to mediate the endocytosis of TFR1, increasing the levels of both cellular total iron and ferrous iron and thereby promoting ferroptosis.
references:
- id: GO_REF:0000002
  title: Gene Ontology annotation through association of InterPro records with GO
    terms
  findings: []
- id: GO_REF:0000003
  title: Gene Ontology annotation based on Enzyme Commission mapping
  findings: []
- id: GO_REF:0000024
  title: Manual transfer of experimentally-verified manual GO annotation data to orthologs
    by curator judgment of sequence similarity
  findings: []
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000044
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location
    vocabulary mapping, accompanied by conservative changes to GO terms applied by
    UniProt
  findings: []
- id: GO_REF:0000052
  title: Gene Ontology annotation based on curation of immunofluorescence data
  findings: []
- id: GO_REF:0000116
  title: Automatic Gene Ontology annotation based on Rhea mapping
  findings: []
- id: PMID:11877457
  title: "Phosphorylation of the AP2 mu subunit by AAK1 mediates high affinity binding to membrane protein sorting signals."
  findings:
  - statement: AAK1 is the endogenous candidate kinase for AP2M1/μ2 Thr156.
    supporting_text: >-
      Together, these data suggest that AAK1 corresponds to the endogenous kinase
      responsible for phosphorylation of μ2 at Thr-156.
    reference_section_type: RESULTS
  - statement: AAK1 phosphorylation of AP2M1/μ2 increases AP2 affinity for cargo sorting signals.
    supporting_text: >-
      Thus, our in vitro experiments demonstrate that AP2 affinity for sorting signals
      can be significantly regulated by AAK1-mediated phosphorylation of μ2.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct site-mapping and biochemical evidence for the core AAK1-AP2M1 mechanism.
      AP2 was purified from pig brain and tested with recombinant AAK1, so this is
      conserved mammalian biochemistry rather than a purely human-cell assay.
- id: PMID:11877461
  title: "Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin-mediated endocytosis."
  findings:
  - statement: AAK1 directly binds the AP2 α-adaptin subunit in vitro.
    supporting_text: Thus, these results demonstrate that AAK1 interacts directly with α-adaptin in vitro.
    reference_section_type: RESULTS
  - statement: AAK1 preferentially phosphorylates μ subunits of AP complexes.
    supporting_text: >-
      These results provide strong evidence that μ is a specific target of AAK1
      phosphorylation in CCVs and suggests that the observed phosphorylation of μ can be
      attributed to the presence of endogenous AAK1 in these samples (Fig. 4).
    reference_section_type: RESULTS
  - statement: AAK1 regulates an early AP2-dependent step of clathrin-mediated endocytosis.
    supporting_text: >-
      Collectively, these data provide functional evidence that AAK1 plays a role in
      AP2-stimulated endocytosis at an early step by specifically phosphorylating the μ
      subunit of the AP complex.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Foundational direct interaction, kinase, localization, and endocytosis evidence.
      The study uses human full-length AAK1 together with rat/bovine preparations and
      human HeLa/A431 cell assays, so individual findings retain their model context.
- id: PMID:12952931
  title: "Differential requirements for AP-2 in clathrin-mediated endocytosis."
  findings:
  - statement: Efficient AP2 recruitment and μ2 phosphorylation require AAK1's α-adaptin-interacting domain.
    supporting_text: >-
      We conclude that efficient recruitment of AAK1 to AP-2 and its phosphorylation of
      μ2 requires the AID and that each of the individual domains and truncated mutants
      retain their expected activities.
    reference_section_type: RESULTS
  - statement: Full-length AAK1 overexpression binds and disrupts AP2 function in human cells.
    supporting_text: These data establish that AAK1 can bind to and disrupt the function of AP-2 complexes in vivo.
    reference_section_type: RESULTS
  - statement: Approximately 80% AAK1 knockdown did not measurably change transferrin uptake, AP2 distribution, or μ2 phosphorylation in A549 or HeLa cells.
    supporting_text: >-
      Transfection of two different siRNAs that specifically target AAK1 reduced AAK1
      expression by ∼80% in either A549 or HeLa cells. However, in neither case did we
      observe an alteration in Tfn internalization, AP-2 distribution, or μ2
      phosphorylation (Fig. S2, available at
      http://www.jcb.org/cgi/content/full/jcb.200304069/DC1).
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct human-cell AP2/endocytosis study, but its positive phenotype is driven by
      full-length AAK1 overexpression and is kinase-independent; partial RNAi produced no
      phenotype, an important redundancy/residual-activity caveat.
- id: PMID:14617351
  title: "AAK1-mediated micro2 phosphorylation is stimulated by assembled clathrin."
  full_text_unavailable: true
  findings:
  - statement: Clathrin stimulates AAK1-dependent AP2 μ2 phosphorylation.
    supporting_text: >-
      Here, we report that AAK1 is an atypical kinase that is rate limited by its stable
      association with AP2 and that clathrin stimulates micro2 phosphorylation by AAK1.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: Assembled clathrin cages stimulate AAK1 more strongly than unassembled triskelia.
    supporting_text: >-
      Importantly, incubation of AAK1 with clathrin cages resulted in even greater
      stimulation when compared to that of unassembled clathrin triskelia.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      The cached abstract directly supports clathrin-dependent activation of AAK1, a
      core regulatory mechanism. Full experimental and species details are unavailable
      locally, so no assay context beyond the abstract is inferred.
- id: PMID:17494869
  title: "A novel AAK1 splice variant functions at multiple steps of the endocytic pathway."
  full_text_unavailable: true
  findings:
  - statement: The long AAK1 isoform has an extended C terminus with a second, directly clathrin-binding domain.
    supporting_text: >-
      Here we have identified a long form of AAK1 (AAK1L) that contains an extended
      C-terminus that encodes an additional clathrin-binding domain (CBD2) consisting of
      multiple low-affinity interaction motifs. Protein interaction studies demonstrate
      that AAK1L CBD2 directly binds clathrin.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: The long and short AAK1 isoforms show similar basal and clathrin-stimulated kinase activity toward AP2 μ2 in vitro.
    supporting_text: >-
      However, in vitro kinase assays reveal little difference between AAK1 isoforms in
      their basal or clathrin-stimulated kinase activity toward the AP-2 micro2 subunit.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: AAK1 depletion impairs transferrin recycling from early/sorting endosomes, supporting an endosomal recycling role in addition to internalization.
    supporting_text: >-
      Surprisingly, CBD2 overexpression or AAK1 depletion by RNA interference
      significantly impairs transferrin recycling from the early/sorting endosome.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct isoform, clathrin-binding, AP2-kinase, and loss-of-function recycling
      evidence. The cache contains only the abstract plus a failed PDF placeholder, so
      detailed cell/species context cannot be independently audited.
- id: PMID:18657069
  title: "AAK1 regulates Numb function at an early step in clathrin-mediated endocytosis."
  full_text_unavailable: true
  findings:
  - statement: The paper reports AAK1-dependent redistribution of Numb between perinuclear endosomes and the plasma membrane; this localization phenotype remains usable with overexpression/depletion caveats.
    supporting_text: >-
      We find that AAK1 redistributes Numb to perinuclear endosomes when overexpressed,
      while kinase depletion causes Numb to accumulate at the plasma membrane.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: The paper reports Numb Thr102 as an AAK1 phosphorylation site, but that substrate/site assignment is disputed by PMID:42082516.
    supporting_text: >-
      Overexpression of a Numb point mutant (T102A) that lacks the AAK1 phosphorylation
      site potently disrupts transferrin and low-density lipoprotein internalization but
      does not impact EGF uptake.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  reference_review:
    relevance: HIGH
    correctness: DISPUTED
    review_notes: >-
      The cached abstract supports the reported Numb localization and cargo-selective
      phenotypes, but the central Numb-T102 substrate claim is contradicted by
      PMID:42082516: purified AAK1/BMP2K did not phosphorylate the site, RPE
      phosphoproteomics did not detect it, and its sequence lacks the refined recognition
      motif. Preserve the localization phenotype; do not treat T102 phosphorylation as
      established.
- id: PMID:21464124
  title: "The adaptor-associated kinase 1, AAK1, is a positive regulator of the Notch pathway."
  findings:
  - statement: AAK1 directly interacts with the membrane-tethered activated form of Notch.
    supporting_text: >-
      We show here that AAK1, the adaptor-associated kinase 1, directly interacts with the
      membrane-tethered active form of Notch released by metalloprotease cleavage.
    reference_section_type: ABSTRACT
  - statement: Active AAK1 stabilizes activated and monoubiquitinated Notch upstream of γ-secretase cleavage.
    supporting_text: >-
      Active AAK1 acts upstream of the γ-secretase cleavage by stabilizing both the
      membrane-tethered activated form of Notch and its monoubiquitinated counterpart.
    reference_section_type: ABSTRACT
  - statement: AAK1 promotes activated Notch localization to Rab5-positive endosomes, whereas AAK1 depletion interferes with that localization.
    supporting_text: >-
      Moreover, transfected AAK1 increases the localization of activated Notch to
      Rab5-positive endocytic vesicles, while AAK1 depletion or overexpression of Numb,
      an inhibitor of the pathway, interferes with this localization.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct mammalian-cell evidence supports activated-Notch binding, stabilization,
      endosomal sorting, and positive pathway regulation. Its secondary Numb model is not
      propagated because that arm depends on the now-disputed PMID:18657069 claim.
- id: PMID:22645275
  title: "Identification of novel ATP13A2 interactors and their role in α-synuclein misfolding and toxicity."
  full_text_unavailable: true
  findings:
  - statement: A membrane yeast-two-hybrid screen reported 43 ATP13A2 interactors, but the cached abstract does not identify AAK1 among them.
    supporting_text: >-
      Utilizing a split-ubiquitin membrane yeast two-hybrid system that was developed to
      identify interacting partners of full-length integral membrane proteins, we
      identified 43 novel interactors that primarily implicate ATP13A2 in cellular
      processes such as endoplasmic reticulum (ER) translocation, ER-to-Golgi trafficking
      and vesicular transport and fusion.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  reference_review:
    relevance: LOW
    correctness: UNVERIFIED
    review_notes: >-
      The cached abstract verifies the ATP13A2 interactor screen but contains no AAK1 or
      AAK1L mention. Because the full text and supplement are unavailable in cache, the
      pair cannot be evaluated mechanistically; the generic protein-binding term remains
      uninformative.
- id: PMID:23781025
  title: "Αvβ3-integrin-mediated adhesion is regulated through an AAK1L- and EHD3-dependent rapid-recycling pathway."
  full_text_unavailable: true
  findings:
  - statement: AAK1L and EHD3 are required for αvβ3-integrin-mediated adhesion in human HeLa cells.
    supporting_text: >-
      We discovered that two early endosome factors, AAK1L and EHD3, are critical for
      αvβ3-integrin-mediated cell adhesion in HeLa cells.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: Depletion of either AAK1L or EHD3 delays short-loop β3-integrin recycling from early endosomes to the cell surface.
    supporting_text: >-
      siRNA-mediated depletion of either factor delays short-loop β3 integrin recycling
      from the early endosome back to the cell surface.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  - statement: AAK1L kinase activity and its C-terminal domain are required to maintain cell adhesion.
    supporting_text: >-
      Moreover, structure-function analysis reveals that AAK1L kinase activity, as well
      as its C-terminal domain, is essential for cell adhesion maintenance.
    reference_section_type: ABSTRACT
    full_text_unavailable: true
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct human HeLa-cell depletion and structure-function evidence for the long AAK1
      isoform. EHD3 is a cooperating factor, so the recycling phenotype is not
      attributable solely to AAK1L.
- id: PMID:26853940
  title: "Family-wide Structural Analysis of Human Numb-Associated Protein Kinases."
  findings:
  - statement: The human AAK1 kinase domain was structurally resolved at 1.95 Å in an inhibitor-bound complex.
    supporting_text: >-
      The structure of AAK1 was solved in complex with a small-molecule inhibitor to
      1.95 Å resolution in space group P212121 with two AAK1 kinase domains per
      asymmetric unit (chains A and B) (Table 1).
    reference_section_type: RESULTS
  - statement: Recombinant AAK1 directly phosphorylated an AP2-medium-subunit-derived peptide in vitro.
    supporting_text: >-
      A 17-mer synthetic peptide corresponding to the AAK1/BIKE phosphorylation site on
      the medium subunit of AP2 was used to test kinase activity. Phosphorylation of the
      substrate in the presence of AAK1 was confirmed by mass spectrometry (Figure S1).
    reference_section_type: RESULTS
  - statement: AAK1's assembled regulatory spine without activation-loop phosphorylation supports constitutive catalytic competence.
    supporting_text: >-
      Similarly, in structures of BIKE and AAK1 there is alignment of the R spine in the
      absence of phosphorylation, suggesting that NAK family members are constitutively
      active kinases (Figure 2C).
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct structural and biochemical characterization of human AAK1. Constitutive
      activity is a kinase-domain structural inference; regulation of full-length AAK1
      in cells remains context dependent.
- id: PMID:30605688
  title: "WNT Activates the AAK1 Kinase to Promote Clathrin-Mediated Endocytosis of LRP6 and Establish a Negative Feedback Loop."
  findings:
  - statement: Human cell-line perturbations establish AAK1 as a negative regulator of WNT signaling across multiple tissue-derived models.
    supporting_text: Together, these data establish that AAK1 negatively regulates WNT signaling in cells derived from multiple tissue types.
    reference_section_type: RESULTS
  - statement: AAK1 promotes clathrin-mediated endocytosis and lysosomal degradation of LRP6, forming a negative-feedback mechanism for WNT signaling.
    supporting_text: >-
      Together, these results suggest that AAK1 inhibits WNT signaling by inducing CME
      endocytosis and lysosomal degradation of LRP6.
    reference_section_type: RESULTS
  - statement: Delayed WNT3A-induced phosphorylation of AP2M1 requires AAK1 expression and kinase activity.
    supporting_text: The WNT3A-induced phosphorylation of AP2M1 required AAK1 expression and activity.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct gain-of-function, knockdown, inhibitor, trafficking, and AP2M1-phosphorylation
      evidence in human HEK293T, HT1080, RKO, and related cell models.
- id: PMID:34315807
  title: "Dysregulation of the AP2M1 phosphorylation cycle by LRRK2 impairs endocytosis and leads to dopaminergic neurodegeneration."
  findings:
  - statement: With equimolar recombinant kinases, AAK1 phosphorylated AP2M1 more potently than LRRK2 in vitro.
    supporting_text: Our results showed that AAK1 was more potent than LRRK2 in phosphorylating AP2M1 in vitro.
    reference_section_type: RESULTS
  - statement: In human SH-SY5Y cells, AAK1 increased endogenous AP2M1 phosphorylation independently of LRRK2.
    supporting_text: >-
      We found that AAK1 increased the amount of endogenous AP2M1 phosphorylation in both
      LRRK2 WT and KO cells (Fig. 3, E and F), suggesting that AAK1 can phosphorylate
      AP2M1 independently of LRRK2.
    reference_section_type: RESULTS
  - statement: Mouse tissue evidence indicates low relative AAK1 abundance in brain but high abundance in thyroid.
    supporting_text: >-
      Notably, we found that unlike LRRK2, the expression of AAK1 was relatively low in
      mouse brain tissue (Fig. 3, I and J). In contrast, AAK1 was highly expressed in the
      mouse thyroid gland where LRRK2 expression was relatively low.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct AP2M1 kinase evidence includes recombinant assays and human SH-SY5Y cells.
      Tissue-distribution conclusions are specifically from mouse and are not presented
      as direct human tissue evidence.
- id: PMID:41407700
  title: "AAK1 activation-mediated iron trafficking drives ferroptotic cell death."
  findings:
  - statement: Recombinant active PKCβII directly phosphorylates AAK1; phosphatase treatment removes the signal.
    supporting_text: >-
      The level of Ser/Thr phosphorylation of AAK1 protein increased significantly in the
      presence of recombinant active PKCβII kinase, but was blocked in the addition of
      λ-phosphatase (Fig. 3E, F).
    reference_section_type: RESULTS
  - statement: Activated AAK1 phosphorylates AP2M1, promoting clathrin-dependent TFR1 endocytosis, cellular iron accumulation, and ferroptosis.
    supporting_text: >-
      Mechanistically, activated AAK1 phosphorylates AP2M1, which facilitates the
      recruitment of clathrin to mediate the endocytosis of TFR1, increasing the levels
      of both cellular total iron and ferrous iron and thereby promoting ferroptosis.
    reference_section_type: ABSTRACT
  - statement: In nude-mouse xenografts of human MDA-MB-231 cells, AAK1 loss reduced IKE-induced ferroptosis and increased tumor growth.
    supporting_text: >-
      To further confirm the role of AAK1 during the process of ferroptosis in vivo, we
      performed xenograft tumor model by inoculating AAK1-knockout MDA-MB-231 cells with
      or without transfecting plasmids of wild-type or S670/T674Ala mutant AAK1 into nude
      mice. These nude mice were treated with IKE (a lipid-soluble form of erastin for
      animal experiment) or DMSO as control. As expected, knockout of AAK1 significantly
      inhibited ferroptosis through resistance of lipid peroxidation induced by IKE and
      promoted tumor growth (Fig. 6G–J).
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct mechanistic evidence in multiple human cancer cell lines identifies the
      PKCβII-AAK1-AP2M1-TFR1 iron-uptake axis. The tumor-growth result is a human-cell
      xenograft in mice, and ferroptosis is an induced contextual role rather than
      necessarily AAK1's basal core function.
- id: PMID:42082516
  title: "AAK1-mediated phosphorylation of PDLIM5 and Talin1 promotes focal adhesion disassembly to accelerate cell migration."
  findings:
  - statement: Biochemical and phosphoproteomic screens identify PDLIM5 and Talin1 as direct AAK1/BMP2K substrates.
    supporting_text: >-
      Here, using motif-guided in silico, biochemical, and phosphoproteomic screens, we
      identify PDLIM5 and Talin1 as direct AAK1/BMP2K substrates.
    reference_section_type: ABSTRACT
  - statement: AAK1 has a specific, nonredundant kinase-dependent role in focal-adhesion turnover and cell migration in human RPE cells.
    supporting_text: >-
      AAK1 deletion or siRNA-mediated knockdown reproducibly impaired RPE migration by
      ~25% (Fig. 3d, e and Supplementary Fig. 1h), whereas BMP2K loss had no measurable
      effect, revealing a specific and non-redundant role for AAK1 in cell motility.
    reference_section_type: RESULTS
  - statement: Human RPE-cell phosphoproteomics and in-vitro kinase assays failed to support NUMB Thr102 as an AAK1/BMP2K substrate, directly disputing the PMID:18657069 claim.
    supporting_text: >-
      Furthermore, our global phosphoproteome profiling of RPE cells failed to detect
      phosphorylation of the previously proposed substrates Dab2 T109, Arh T104, or NUMB
      T102. This conclusion is supported by both our in vitro kinase assays and the
      absence of phosphorylation evidence in high-throughput databases, such as
      PhosphoSitePlus (Fig. 1h, i), which suggests that Dab2 T109, Arh T104, and NUMB T102
      are unlikely to be physiological targets of AAK1 or BMP2K.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: >-
      Direct human RPE-cell genetics, biochemistry, phosphoproteomics, imaging, and
      focal-adhesion assays. The PDLIM5/Talin1 discovery initially combines AAK1 and
      BMP2K perturbation, but AAK1-specific genetics establish the migration role. Its
      negative NUMB Thr102 evidence materially disputes PMID:18657069 and is recorded as
      counterevidence rather than silently accepting the older claim.
- id: Reactome:R-HSA-199991
  title: Membrane Trafficking
  findings: []
- id: Reactome:R-HSA-8856808
  title: Recruitment of AP-2 complex and clathrin
  findings: []
- id: Reactome:R-HSA-8856813
  title: AAK1 phosphorylates AP-2 mu subunit at T156
  findings: []
- id: Reactome:R-HSA-8866283
  title: ARBB recruits GPCRs into clathrin-coated pits
  findings: []
- id: Reactome:R-HSA-8867754
  title: F- and N- BAR domain proteins bind the clathrin-coated pit
  findings: []
- id: Reactome:R-HSA-8867756
  title: CLASP proteins and cargo are recruited to the nascent clathrin-coated pit
  findings: []
- id: Reactome:R-HSA-8868071
  title: Clathrin recruits PIK3C2A
  findings: []
- id: Reactome:R-HSA-8868072
  title: Clathrin-associated PIK3C2A phosphorylates PI(4)P to PI(3,4)P2
  findings: []
- id: Reactome:R-HSA-8868230
  title: SNX9 recruits components of the actin polymerizing machinery
  findings: []
- id: Reactome:R-HSA-8868236
  title: BAR domain proteins recruit dynamin
  findings: []
- id: Reactome:R-HSA-8868648
  title: SYNJ hydrolyze PI(4,5)P2 to PI(4)P
  findings: []
- id: Reactome:R-HSA-8868651
  title: Endophilins recruit synaptojanins to the clathrin-coated pit
  findings: []
- id: Reactome:R-HSA-8868658
  title: HSPA8-mediated ATP hydrolysis promotes vesicle uncoating
  findings: []
- id: Reactome:R-HSA-8868659
  title: Clathrin recruits auxilins to the clathrin-coated vesicle
  findings: []
- id: Reactome:R-HSA-8868660
  title: Auxilin recruits HSPA8:ATP to the clathrin-coated vesicle
  findings: []
- id: Reactome:R-HSA-8868661
  title: Dynamin-mediated GTP hydrolysis promotes vesicle scission
  findings: []
- id: Reactome:R-HSA-8871193
  title: Dissociation of AAK1 and dephosphorylation of AP-2 mu2
  findings: []
- id: Reactome:R-HSA-8871194
  title: RAB5 and GAPVD1 bind AP-2
  findings: []
core_functions:
- description: >-
    At plasma-membrane clathrin-coated pits, AAK1 binds AP-2 and clathrin and
    uses clathrin-stimulated protein serine/threonine kinase activity to
    phosphorylate AP2M1/mu2 at Thr-156. This increases AP-2 cargo-sorting
    affinity and regulates clathrin-dependent cargo internalization; AAK1 also
    positively regulates return of internalized cargo from early/sorting
    endosomes to the cell surface.
  molecular_function:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  directly_involved_in:
  - id: GO:2000369
    label: regulation of clathrin-dependent endocytosis
  - id: GO:2001137
    label: positive regulation of endocytic recycling
  locations:
  - id: GO:0005905
    label: clathrin-coated pit
  substrates:
  - id: UniProtKB:Q96CW1
    label: AP-2 complex subunit mu
  supported_by:
  - reference_id: PMID:11877457
    supporting_text: >-
      Together, these data suggest that AAK1 corresponds to the endogenous kinase responsible for phosphorylation of μ2 at Thr-156.
  - reference_id: PMID:14617351
    supporting_text: >-
      Importantly, incubation of AAK1 with clathrin cages resulted in even greater stimulation when compared to that of unassembled clathrin triskelia.
  - reference_id: PMID:17494869
    supporting_text: >-
      Surprisingly, CBD2 overexpression or AAK1 depletion by RNA interference significantly impairs transferrin recycling from the early/sorting endosome.
  - reference_id: PMID:34315807
    supporting_text: >-
      We found that AAK1 increased the amount of endogenous AP2M1 phosphorylation in both LRRK2 WT and KO cells (Fig. 3, E and F), suggesting that AAK1 can phosphorylate AP2M1 independently of LRRK2.
- description: >-
    At mature focal adhesions, AAK1 kinase activity promotes timely release of
    adhesome components during focal-adhesion disassembly, increasing adhesion
    turnover and cell migration. PDLIM5 Thr-290 and talin-1 Thr-2270 are direct
    in-vitro substrates of both AAK1 and BMP2K and are jointly kinase-responsive
    in cells; the individual endogenous contribution of AAK1 to each phosphosite
    remains unresolved.
  molecular_function:
    id: GO:0004674
    label: protein serine/threonine kinase activity
  directly_involved_in:
  - id: GO:0120183
    label: positive regulation of focal adhesion disassembly
  - id: GO:0030335
    label: positive regulation of cell migration
  locations:
  - id: GO:0005925
    label: focal adhesion
  substrates:
  - id: UniProtKB:Q96HC4
    label: PDZ and LIM domain protein 5
  - id: UniProtKB:Q9Y490
    label: Talin-1
  supported_by:
  - reference_id: PMID:42082516
    supporting_text: >-
      Here, using motif-guided in silico, biochemical, and phosphoproteomic screens, we identify PDLIM5 and Talin1 as direct AAK1/BMP2K substrates.
  - reference_id: PMID:42082516
    supporting_text: >-
      Because rapid paxillin turnover is essential for efficient cell migration, these data position AAK1 as a positive regulator of FA disassembly and motility.
  - reference_id: PMID:42082516
    supporting_text: >-
      Thus, AAK1 is specifically recruited to mature, tension-bearing FAs and thereby positioned to efficiently phosphorylate substrates PDLIM5 and Talin1.
knowledge_gaps:
- gap_statement: >-
    The cargo- and cell-type-specific division of AP2M1 phosphorylation and
    clathrin-dependent endocytosis among AAK1, BMP2K, GAK, and other kinases is
    not resolved.
  boundary: >-
    AAK1 directly phosphorylates AP2M1 and regulates endocytosis and recycling in
    multiple systems, but in human RPE cells BMP2K is the dominant NAK for
    receptor internalization while AAK1 loss has only a modest effect on AP2M1
    phosphorylation.
  gap_kind:
  - BIOLOGY
  dark_aspect: RESIDUAL_SUBGAP
  status: NARROWING
  significance: >-
    Resolving the division of labor is necessary to predict which cargos and
    tissues will respond to selective AAK1 perturbation and to avoid treating
    all NAK-family endocytic evidence as interchangeable.
  resolution: >-
    Parallel endogenous single- and combinatorial-kinase perturbations, coupled
    to AP2M1 phosphosite quantification and cargo-resolved uptake and recycling
    assays across cell types, would define nonredundant and compensatory roles.
  provenance:
  - reference_id: PMID:42082516
    supporting_text: >-
      Together, these data indicate that BMP2K, rather than AAK1, is the primary NAK regulating CME in RPE cells.
- gap_statement: >-
    The individual contribution of AAK1 versus BMP2K to endogenous PDLIM5
    Thr-290 and talin-1 Thr-2270 phosphorylation in intact cells remains
    unresolved.
  boundary: >-
    Recombinant kinase assays and double-knockout phosphoproteomics establish
    both sites as AAK1/BMP2K-responsive, while AAK1-specific loss and rescue
    establish the focal-adhesion turnover and migration phenotype.
  gap_kind:
  - BIOLOGY
  dark_aspect: RESIDUAL_SUBGAP
  status: NARROWING
  significance: >-
    Site-specific kinase attribution would close the causal link between AAK1's
    focal-adhesion recruitment, its two proposed adhesome substrates, and the
    nonredundant migration phenotype.
  resolution: >-
    Quantify both phosphosites in matched AAK1 and BMP2K single knockouts and
    acute degron lines, followed by wild-type, kinase-dead, and
    recruitment-defective rescue and focal-adhesion lifetime measurements.
  provenance:
  - reference_id: PMID:42082516
    supporting_text: >-
      The use of double knock-out cells allowed us to capture phosphorylation driven by either kinase while mitigating potential paralogue redundancy.
proposed_new_terms: []
suggested_questions:
- question: >-
    Which cargo classes and human cell types depend nonredundantly on AAK1,
    rather than BMP2K, GAK, or another kinase, for AP2M1 phosphorylation,
    internalization, and endocytic recycling?
- question: >-
    Do AAK1L and AAK1S differ in recruitment to coated pits, early endosomes,
    and focal adhesions, or in their relative contributions to recycling and
    adhesion turnover?
suggested_experiments:
- hypothesis: >-
    AAK1 and BMP2K make cell- and cargo-specific, partly compensatory
    contributions to AP2M1 phosphorylation and endocytic flux.
  description: >-
    Generate matched endogenous AAK1, BMP2K, GAK, and combinatorial acute-degron
    cell panels in several human lineages. Measure AP2M1 Thr-156 phosphorylation,
    coated-pit dynamics, and uptake/recycling kinetics for transferrin receptor,
    integrins, LRP6, and TFR1, with wild-type and kinase-dead rescues.
  experiment_type: >-
    Acute genetic perturbation, targeted phosphoproteomics, and quantitative
    live-cell endocytic trafficking
- hypothesis: >-
    AAK1 isoforms and C-terminal recruitment motifs partition its endocytic and
    focal-adhesion activities.
  description: >-
    Use isoform-selective CRISPR editing and endogenous fluorescent tagging to
    compare AAK1L and AAK1S recruitment at coated pits, early endosomes, and
    focal adhesions. Combine isoform-specific rescue with AP2M1, PDLIM5, and
    talin-1 phosphosite measurements, cargo recycling, focal-adhesion lifetime,
    and migration assays.
  experiment_type: >-
    Isoform-resolved genome editing, live-cell imaging, and targeted
    phosphoproteomics

Gene Description

Existing Annotations Review

Core Functions

References

Suggested Questions for Experts

Suggested Experiments

Knowledge Gaps

📚 Additional Documentation

Notes

AAK1 review notes

Research provenance

Established endocytic mechanism

Endocytic scope and boundaries

Revised substrate scope and focal adhesions

Context-specific signaling and stress outputs

Evidence and annotation boundaries

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