GRAP2 (GADS, also called GrpL, Grf40, Mona) is a hematopoietic-restricted cytosolic adaptor of the GRB2 family with no catalytic activity. It has an N-terminal SH3 domain, a central SH2 domain, a unique ~120-residue glutamine/proline-rich linker and a C-terminal SH3 domain. The C-terminal SH3 domain binds an atypical, non-PxxP R-x-x-K motif in the scaffold SLP-76 (LCP2) with nanomolar affinity, so GADS and SLP-76 form a constitutive complex. After antigen-receptor engagement, the GADS SH2 domain docks on ZAP70/SYK-phosphorylated LAT (pY171 and pY191 in the conventional short-isoform numbering; pY200 and pY220 in canonical LAT O43561-1), recruiting SLP-76 to the LAT signalosome at the plasma membrane, where SLP-76 assembles PLCG1, ITK, VAV1 and NCK. GADS thereby couples the T cell receptor to PLCG1 phosphorylation, calcium flux and NFAT activation, sets the sensitivity of T cells to weak TCR stimuli, and acts the same way downstream of FcERI in mast cells and of GPVI/CLEC-2 in platelets (where it is partly redundant with GRB2). Its activity is tuned by dimerization, ITK phosphorylation of Y45, phosphorylation of T262 and caspase cleavage of the linker. The C-terminal SH3 domain also binds HPK1 (MAP4K1), and the SH2 domain binds phosphorylated SHC, KIT, the M-CSF receptor, LAX1 and CD6. GADS-deficient mice have impaired thymocyte proliferation and selection and severely impaired IgE-mediated mast cell activation.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0001784 phosphotyrosine residue binding | IBA GO_REF:0000033 | ACCEPT | Summary: IBA phosphotyrosine residue binding (GRB2-family node). Reason: The GADS SH2 domain binds phosphotyrosine motifs (pY-x-N-x) on LAT, LAX, SHC and receptor tyrosine kinases; crystal structures and calorimetry show selective binding to LAT pY171/pY191 (short-isoform numbering; pY200/pY220 in canonical LAT O43561-1). Inherited GRB2-family SH2 activity, correctly placed. Supporting Evidence: PMID:15029250 Gads-SH2 binds LAT tyrosine phosphorylation sites 171 and 191 with higher affinities than the other sites PMID:10021361 this interaction was mediated by the Gads SH2 domain |
| GO:0005515 protein binding | IPI PMID:12359715 Molecular cloning of a novel gene encoding a membrane-associ... | MODIFY | Summary: IPI protein binding with the transmembrane adaptor LAX1 (PMID:12359715). Reason: LAX, like LAT, is tyrosine-phosphorylated after antigen-receptor stimulation and then binds GADS; this is the GADS SH2 domain recognising phosphotyrosine motifs. Replace protein binding with phosphotyrosine residue binding (abstract-only paper; the abstract states the tyrosine-motif basis). Proposed replacements: phosphotyrosine residue binding Supporting Evidence: PMID:12359715 LAX has tyrosine motifs for binding Grb2, Gads, and the p85 subunit of phosphatidylinositol 3-kinase |
| GO:0005515 protein binding | IPI PMID:15929943 Novel Src homology 3 domain-binding motifs identified from p... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a SH3-domain peptide-array screen of the SLP-76 proline-rich region (PMID:15929943). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:16189514 Towards a proteome-scale map of the human protein-protein in... | REMOVE | Summary: IPI protein binding with STAMBP from a CCSB-HI1 yeast two-hybrid screen (PMID:16189514). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:16189514 Towards a proteome-scale map of the human protein-protein in... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a CCSB-HI1 yeast two-hybrid screen (PMID:16189514). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:16273093 A quantitative protein interaction network for the ErbB rece... | MODIFY | Summary: IPI protein binding with ERBB2 from a protein-microarray screen of SH2/PTB domains against ErbB phosphopeptides (PMID:16273093). Reason: The assay measured binding of isolated SH2 domains to ErbB phosphotyrosine peptides, so the informative activity captured is phosphotyrosine residue binding by the GADS SH2 domain. ERBB2 is not a physiological GADS partner in leukocytes; the replacement records only the domain activity. Proposed replacements: phosphotyrosine residue binding Supporting Evidence: PMID:16273093 we use protein microarrays comprising virtually every Src homology 2 (SH2) and phosphotyrosine binding (PTB) domain encoded in the human genome |
| GO:0005515 protein binding | IPI PMID:16730941 A systematic analysis of human CHMP protein interactions: ad... | REMOVE | Summary: IPI protein binding with STAMBP from a CHMP interaction screen (PMID:16730941). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:17010654 Crystal structure of the C-terminal SH3 domain of the adapto... | MODIFY | Summary: IPI protein binding with SLP-76 (LCP2) from PMID:17010654, a mechanistic study of the GADS C-terminal SH3 domain / SLP-76 RxxK interaction. Reason: The interaction is real and central to GRAP2 function, but "protein binding" is uninformative. The C-SH3 of GADS constitutively binds the SLP-76 RxxK motif and its SH2 binds phospho-LAT, so the physiological activity is bridging SLP-76 to LAT, i.e. signaling adaptor activity (the same MF used for LCP2 in this project). Proposed replacements: signaling adaptor activity Supporting Evidence: PMID:17010654 Following T cell receptor ligation, GADS interacts through its C-terminal SH3 domain with the adaptors SLP-76 and LAT, to form a multiprotein signaling complex that is crucial for T cell activation PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005515 protein binding | IPI PMID:17065230 A noncanonical SH3 domain binding motif links BK channels to... | REMOVE | Summary: IPI protein binding with mouse Kcnma1 from a BK channel SH3-binding screen (PMID:17065230). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:17235283 Efficient T-cell receptor signaling requires a high-affinity... | MODIFY | Summary: IPI protein binding with SLP-76 (LCP2) from PMID:17235283, a mechanistic study of the GADS C-terminal SH3 domain / SLP-76 RxxK interaction. Reason: The interaction is real and central to GRAP2 function, but "protein binding" is uninformative. The C-SH3 of GADS constitutively binds the SLP-76 RxxK motif and its SH2 binds phospho-LAT, so the physiological activity is bridging SLP-76 to LAT, i.e. signaling adaptor activity (the same MF used for LCP2 in this project). Proposed replacements: signaling adaptor activity Supporting Evidence: PMID:17235283 In signaling from the T-cell receptor (TCR), the Gads C-terminal SH3 domain binds a core RxxK sequence motif in the SLP-76 scaffold PMID:17235283 Gads, on the other hand, links phosphorylated LAT to the scaffold protein, SLP-76, which binds the Gads C-SH3 domain PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005515 protein binding | IPI PMID:20534575 SH2 domain containing leukocyte phosphoprotein of 76-kDa (SL... | MODIFY | Summary: IPI protein binding with SLP-76 (LCP2) from PMID:20534575, a mechanistic study of the GADS C-terminal SH3 domain / SLP-76 RxxK interaction. Reason: The interaction is real and central to GRAP2 function, but "protein binding" is uninformative. The C-SH3 of GADS constitutively binds the SLP-76 RxxK motif and its SH2 binds phospho-LAT, so the physiological activity is bridging SLP-76 to LAT, i.e. signaling adaptor activity (the same MF used for LCP2 in this project). Proposed replacements: signaling adaptor activity Supporting Evidence: PMID:20534575 ZAP-70 phosphorylation of LAT that is followed by LAT recruitment of the GADS-SLP-76 complex PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005515 protein binding | IPI PMID:21179510 Histidine domain-protein tyrosine phosphatase interacts with... | REMOVE | Summary: IPI protein binding with PTPN23 (HD-PTP) by yeast two-hybrid, GST pulldown and co-immunoprecipitation (PMID:21179510). Reason: The GrpL C-terminal SH3 domain binds the histidine domain of HD-PTP in overexpression and in vitro assays, but no physiological function of the interaction for GRAP2 has been shown and no informative MF term follows from it. Removing the generic protein binding row does not dispute the interaction. Supporting Evidence: PMID:21179510 another to the C-terminal SH3 domain (amino acid residues 254β330) of Grb2-related protein of the lymphoid system (GrpL) |
| GO:0005515 protein binding | IPI PMID:21516116 Next-generation sequencing to generate interactome datasets. | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a Stitch-seq yeast two-hybrid screen (PMID:21516116). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:24658140 The mammalian-membrane two-hybrid assay (MaMTH) for probing ... | REMOVE | Summary: IPI protein binding with EGFR from a MaMTH membrane two-hybrid screen (PMID:24658140). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:24728074 Enhanced prediction of Src homology 2 (SH2) domain binding p... | MODIFY | Summary: IPI protein binding with KIT from a fluorescence-polarization SH2-domain / phosphopeptide interactome (PMID:24728074). Reason: The assay measured GRAP2 SH2-domain binding to tyrosine-phosphorylated peptides (e.g. KIT pY703/pY936), so the informative activity is phosphotyrosine residue binding. KIT binding by the GADS SH2 domain is also reported independently (PMID:9872323). Proposed replacements: phosphotyrosine residue binding Supporting Evidence: PMID:24728074 Peptides derived from Tyr(P)-703 and Tyr(P)-936 were the predicted GRB2/GRAP2-binding sites based on motif prediction software PMID:9872323 The binding specificity of the Gads SH2 domain is similar to Grb2 and mediates the interaction of Gads with Shc, Bcr-Abl and c-kit |
| GO:0005515 protein binding | IPI PMID:24728074 Enhanced prediction of Src homology 2 (SH2) domain binding p... | MODIFY | Summary: IPI protein binding with GAB1 from a fluorescence-polarization SH2-domain / phosphopeptide interactome (PMID:24728074). Reason: The assay measured GRAP2 SH2-domain binding to tyrosine-phosphorylated peptides (e.g. KIT pY703/pY936), so the informative activity is phosphotyrosine residue binding. KIT binding by the GADS SH2 domain is also reported independently (PMID:9872323). Proposed replacements: phosphotyrosine residue binding Supporting Evidence: PMID:24728074 Peptides derived from Tyr(P)-703 and Tyr(P)-936 were the predicted GRB2/GRAP2-binding sites based on motif prediction software PMID:9872323 The binding specificity of the Gads SH2 domain is similar to Grb2 and mediates the interaction of Gads with Shc, Bcr-Abl and c-kit |
| GO:0005515 protein binding | IPI PMID:25036101 Identification of SH3 domain proteins interacting with the c... | REMOVE | Summary: IPI protein binding with ADAM10 from a SH3-domain pulldown screen of the ADAM10 tail (PMID:25036101). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:25241761 Using an in situ proximity ligation assay to systematically ... | REMOVE | Summary: IPI protein binding with ERBB2 from a proximity-ligation survey in HeLa cells (PMID:25241761). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:25416956 A proteome-scale map of the human interactome network. | REMOVE | Summary: IPI protein binding with SPRY2 from a HI-II-14 yeast two-hybrid screen (PMID:25416956). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:25416956 A proteome-scale map of the human interactome network. | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a HI-II-14 yeast two-hybrid screen (PMID:25416956). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:25814554 Phospho-tyrosine dependent protein-protein interaction netwo... | REMOVE | Summary: IPI protein binding with GAREM1 from a phosphotyrosine-dependent yeast two-hybrid screen (PMID:25814554). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:27107012 Pooled-matrix protein interaction screens using Barcode Fusi... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a Barcode Fusion Genetics yeast two-hybrid screen (PMID:27107012). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:27107012 Pooled-matrix protein interaction screens using Barcode Fusi... | REMOVE | Summary: IPI protein binding with LNX1 from a Barcode Fusion Genetics yeast two-hybrid screen (PMID:27107012). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:28514442 Architecture of the human interactome defines protein commun... | REMOVE | Summary: IPI protein binding with STAMBP from a BioPlex affinity-purification mass spectrometry (PMID:28514442). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:28514442 Architecture of the human interactome defines protein commun... | REMOVE | Summary: IPI protein binding with PTPN23 (HD-PTP) from a BioPlex affinity-purification mass spectrometry (PMID:28514442). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:28514442 Architecture of the human interactome defines protein commun... | REMOVE | Summary: IPI protein binding with ZSCAN21 from a BioPlex affinity-purification mass spectrometry (PMID:28514442). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:29997244 LuTHy: a double-readout bioluminescence-based two-hybrid tec... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a LuTHy two-hybrid benchmark (PMID:29997244). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:31467278 Maximizing binary interactome mapping with a minimal number ... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a binary interactome assay benchmark (PMID:31467278). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:31515488 Extensive disruption of protein interactions by genetic vari... | REMOVE | Summary: IPI protein binding with STAMBP from a variant interaction-perturbation screen (PMID:31515488). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:31980649 Extensive rewiring of the EGFR network in colorectal cancer ... | REMOVE | Summary: IPI protein binding with EGFR from a EGFR-network AP-MS in colorectal cancer cells (PMID:31980649). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with MOS from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with PRR35 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with ZNF250 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with PBLD from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with PRKAA2 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with KPRP from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with Q7Z783 (uncharacterized) from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with RAVER1 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with GAREM1 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with SH2D4A from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with ZNF319 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with GAB2 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: IPI protein binding with ZSCAN21 from a HuRI yeast two-hybrid reference interactome (PMID:32296183). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:32814053 Interactome Mapping Provides a Network of Neurodegenerative ... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a neurodegeneration yeast two-hybrid interactome (PMID:32814053). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with STAMBP from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with GAB1 from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with PTPN23 (HD-PTP) from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with GAREM1 from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with GAB2 from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling... | REMOVE | Summary: IPI protein binding with ZSCAN21 from a BioPlex 3.0 AP-MS (PMID:33961781). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. |
| GO:0005515 protein binding | IPI PMID:37398436 AI-guided pipeline for protein-protein interaction drug disc... | REMOVE | Summary: IPI protein binding with LCP2 (SLP-76) from a PPI-drug-discovery pipeline benchmark set (PMID:37398436). Reason: Generic protein binding from a high-throughput or screening assay; it carries no functional information and the paper does not establish a specific GRAP2 activity with this partner. The GADS-SLP-76 interaction itself is well established and is captured by signaling adaptor activity on the mechanistic rows (PMID:17235283, PMID:17010654, PMID:20534575). |
| GO:0005634 nucleus | IDA PMID:21179510 Histidine domain-protein tyrosine phosphatase interacts with... | KEEP AS NON CORE | Summary: IDA nucleus (PMID:21179510). Reason: GrpL was seen dispersed through the cytosol and nucleus when overexpressed in HEK293T cells (PMID:21179510); there is no evidence that GADS acts in the nucleus, and its function is at the plasma membrane with phospho-LAT. Retained as an observed, non-core location. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus |
| GO:0005634 nucleus | IEA GO_REF:0000044 | KEEP AS NON CORE | Summary: IEA nucleus (GO_REF:0000044). Reason: GrpL was seen dispersed through the cytosol and nucleus when overexpressed in HEK293T cells (PMID:21179510); there is no evidence that GADS acts in the nucleus, and its function is at the plasma membrane with phospho-LAT. Retained as an observed, non-core location. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus |
| GO:0005654 nucleoplasm | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA is_active_in nucleoplasm (GRB2-family node). Reason: GRB2-family node inference; a nuclear pool of GADS is observed (overexpression imaging, PMID:21179510; Human Protein Atlas), but no nuclear function is known and its adaptor activity is exercised at phospho-LAT in the plasma membrane. Kept as a non-core location. |
| GO:0005654 nucleoplasm | IDA GO_REF:0000052 | KEEP AS NON CORE | Summary: IDA nucleoplasm from Human Protein Atlas immunofluorescence (GO_REF:0000052). Reason: Consistent with the nuclear pool seen in overexpression (PMID:21179510); an observed location without a known function. |
| GO:0005737 cytoplasm | IBA GO_REF:0000033 | ACCEPT | Summary: IBA cytoplasm (GO_REF:0000033). Reason: GADS is a soluble cytoplasmic adaptor that is constitutively bound to SLP-76 in the cytosol and recruited to phospho-LAT at the membrane after receptor stimulation. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005737 cytoplasm | IDA PMID:21179510 Histidine domain-protein tyrosine phosphatase interacts with... | ACCEPT | Summary: IDA cytoplasm (PMID:21179510). Reason: GADS is a soluble cytoplasmic adaptor that is constitutively bound to SLP-76 in the cytosol and recruited to phospho-LAT at the membrane after receptor stimulation. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005737 cytoplasm | IEA GO_REF:0000044 | ACCEPT | Summary: IEA cytoplasm (GO_REF:0000044). Reason: GADS is a soluble cytoplasmic adaptor that is constitutively bound to SLP-76 in the cytosol and recruited to phospho-LAT at the membrane after receptor stimulation. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0005737 cytoplasm | TAS PMID:9878555 Molecular cloning and expression of human grap-2, a novel le... | ACCEPT | Summary: TAS cytoplasm (PMID:9878555). Reason: GADS is a soluble cytoplasmic adaptor that is constitutively bound to SLP-76 in the cytosol and recruited to phospho-LAT at the membrane after receptor stimulation. Supporting Evidence: PMID:9878555 The SH2- and SH3-containing adaptor molecules serve to recruit cytosolic signal-transducing molecules to the activated receptor tyrosine kinases |
| GO:0005768 endosome | IDA PMID:21179510 Histidine domain-protein tyrosine phosphatase interacts with... | KEEP AS NON CORE | Summary: IDA endosome (PMID:21179510). Reason: GrpL relocated to endosomes only when co-expressed with HD-PTP constructs carrying the endosome-targeting V-domain; endogenous GADS has not been shown to act on endosomes. Kept as a non-core, partner-dependent location. Supporting Evidence: PMID:21179510 led to the relocation of most of the cytosolic Grb2 and GrpL to endosomes |
| GO:0005768 endosome | IEA GO_REF:0000044 | KEEP AS NON CORE | Summary: IEA endosome (GO_REF:0000044). Reason: GrpL relocated to endosomes only when co-expressed with HD-PTP constructs carrying the endosome-targeting V-domain; endogenous GADS has not been shown to act on endosomes. Kept as a non-core, partner-dependent location. Supporting Evidence: PMID:21179510 led to the relocation of most of the cytosolic Grb2 and GrpL to endosomes |
| GO:0005829 cytosol | IDA GO_REF:0000052 | ACCEPT | Summary: IDA cytosol (GO_REF:0000052). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus |
| GO:0005829 cytosol | IEA GO_REF:0000117 | ACCEPT | Summary: IEA cytosol (GO_REF:0000117). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. Supporting Evidence: PMID:21179510 the rest was dispersed throughout the cytosol and in the nucleus |
| GO:0005829 cytosol | TAS Reactome:R-HSA-1433501 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-1433501). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202216 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202216). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202241 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202241). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202248 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202248). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202325 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202325). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202331 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202331). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-202375 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-202375). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-213407 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-213407). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2396561 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2396561). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2396594 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2396594). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2396606 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2396606). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424476 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424476). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424481 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424481). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424484 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424484). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424485 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424485). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424486 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424486). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2424487 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2424487). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730833 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730833). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730840 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730840). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730841 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730841). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730847 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730847). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730851 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730851). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730856 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730856). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730858 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730858). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730889 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730889). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-2730892 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-2730892). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-389381 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-389381). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005829 cytosol | TAS Reactome:R-HSA-9706304 | ACCEPT | Summary: TAS cytosol (Reactome:R-HSA-9706304). Reason: GADS is a cytosolic adaptor; Reactome places the GADS:SLP-76 complex in the cytosol before its recruitment to phospho-LAT. |
| GO:0005886 plasma membrane | IBA GO_REF:0000033 | ACCEPT | Summary: IBA is_active_in plasma membrane (GRB2-family node). Reason: GADS exercises its adaptor activity at the plasma membrane, where its SH2 domain docks on phospho-LAT and brings SLP-76 into LAT microclusters. Supporting Evidence: PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0007165 signal transduction | IBA GO_REF:0000033 | ACCEPT | Summary: IBA signal transduction. Reason: Correct general parent of the antigen-receptor signaling pathways in which GADS acts as an adaptor; the specific pathways are proposed below. Supporting Evidence: PMID:10021361 Gads may promote cross-talk between the LAT and SLP-76 signaling complexes, thereby coupling membrane-proximal events to downstream signaling pathways |
| GO:0007265 Ras protein signal transduction | TAS PMID:9878555 Molecular cloning and expression of human grap-2, a novel le... | MARK AS OVER ANNOTATED | Summary: TAS Ras protein signal transduction (PMID:9878555). Reason: Unlike GRB2, GADS does not bind SOS; its SH3 domains select SLP-76 and HPK1, and it couples tyrosine-phosphorylated proteins to effectors distinct from SOS and Ras. The cloning paper only speculates about receptor tyrosine kinase signaling. Any effect on Ras/ERK in T cells is indirect, via SLP-76/PLCG1. Supporting Evidence: PMID:9872323 Gads may function to couple tyrosine-phosphorylated proteins such as Shc, Bcr-Abl and activated receptor tyrosine kinases to downstream effectors distinct from Sos and Ras PMID:10209041 GrpL can be coimmunoprecipitated with SLP-76 but not with Sos1 or Sos2 |
| GO:0007267 cell-cell signaling | TAS PMID:9878555 Molecular cloning and expression of human grap-2, a novel le... | MARK AS OVER ANNOTATED | Summary: TAS cell-cell signaling (PMID:9878555). Reason: GADS is an intracellular adaptor acting within receptor signaling in the responding cell; the cloning paper does not support a role in cell-cell signaling as such, and the informative process is the specific receptor signaling pathway (T cell receptor signaling pathway, proposed as NEW). Supporting Evidence: PMID:9878555 Grap-2 may participate in leukocyte-specific protein tyrosine kinase signaling |
| GO:0043408 regulation of MAPK cascade | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA regulation of MAPK cascade (GRB2-family node). Reason: Inherited from GRB2-family adaptors. For GADS there is supporting evidence that it promotes HPK1-mediated JNK activation and prolonged MAPK activation after M-CSF stimulation in myeloid progenitors, but these are secondary to its adaptor role in antigen-receptor signaling. Supporting Evidence: PMID:11313918 Coexpression of Grap2 with HPK1 not only increased the kinase activity of HPK1 in the cell, but also had an additive effect on HPK1 mediated JNK activation |
| GO:0050852 T cell receptor signaling pathway | IMP PMID:25452106 Modulation of TCR responsiveness by the Grb2-family adaptor,... | NEW | Summary: Proposed NEW: GADS is the adaptor that bridges SLP-76 to phospho-LAT in TCR signaling. Reason: Participation test: GADS does part of the work of the pathway, physically linking the SLP-76 scaffold (via its C-SH3/RxxK interaction) to ZAP70-phosphorylated LAT (via its SH2), which assembles the PLCG1-activating signalosome. Comparator check: LAT and LCP2, the other two adaptors of the same trimeric complex, carry this term (reviews in this repository). Evidence: Gads-deficient thymocytes uncouple SLP-76 from LAT and fail to respond to CD3 cross-linking; Gads-deficient Jurkat cells lose sensitivity to weak TCR stimuli. No GOA row currently places GRAP2 in this pathway. Human loss-of-function data agree: GADS-suppressed HuT78 CD4+ T cells show reduced recruitment of SLP-76 and PLCG1 to LAT, calcium flux and cytokine release (PMID:25636200), while GADS-knockout Jurkat cells show network rewiring with largely preserved proximal events but reduced activation and cytokine secretion (PMID:37006259). Supporting Evidence: PMID:25452106 Gads conferred responsiveness to weak TCR stimuli, leading to PLC-Ξ³1 phosphorylation and calcium flux PMID:11239162 Immunoprecipitation experiments revealed that the association between SLP-76 and LAT was uncoupled in GADS- thymocytes PMID:11239162 These observations indicate that GADS is a critical adaptor for CD3 signaling PMID:17235283 Gads, on the other hand, links phosphorylated LAT to the scaffold protein, SLP-76, which binds the Gads C-SH3 domain PMID:25636200 The defect in cytokine production occurred because of impaired calcium mobilization due to reduced recruitment of SLP-76 and PLC-Ξ³1 to the LAT complex |
| GO:0038095 Fc-epsilon receptor signaling pathway | IMP PMID:18664516 Selective impairment of FcepsilonRI-mediated allergic reacti... | NEW | Summary: Proposed NEW: GADS acts as the LAT-SLP-76 bridging adaptor downstream of FcERI in mast cells. Reason: Participation test: in mast cells GADS performs the same bridging adaptor work as in T cells, recruiting SLP-76 to phospho-LAT after FcERI cross-linking. Comparator check: LAT and LCP2 carry this term (reviews in this repository). Evidence: Gads-/- bone marrow-derived mast cells fail to mobilize calcium, degranulate or produce cytokines after FcERI cross-linking, and passive cutaneous anaphylaxis is impaired; disrupting the SLP-76/Gads interaction blocks SLP-76 clustering and FcERI responses. Supporting Evidence: PMID:18664516 Bone marrow-derived mast cells from Gads(-/-) mice failed to induce Ca(2+) mobilization, degranulation and cytokine production upon cross-linking of FcepsilonRI PMID:16479002 the Gads-binding region of SLP-76 regulates clustering of SLP-76 and FcepsilonRI-induced mast cell responses |
| GO:0036398 TCR signalosome | IDA PMID:10021361 The hematopoietic-specific adaptor protein gads functions in... | NEW | Summary: Proposed NEW: GADS is a component of the LAT-GADS-SLP-76 TCR signalosome. Reason: GADS is one of the three adaptors of the heterotrimeric LAT-GADS-SLP-76 complex nucleated on phospho-LAT after TCR engagement; it co-immunoprecipitates phospho-LAT after TCR cross-linking via its SH2 domain while constitutively holding SLP-76. LAT and LCP2 are annotated part_of this complex. Supporting Evidence: PMID:10021361 Gads also coimmunoprecipitated the tyrosine-phosphorylated form of the linker for activated T cells (LAT) adaptor protein following cross-linking of the T-cell receptor PMID:31402911 Three adaptors, LAT, Gads, and SLP-76, form a heterotrimeric complex that mediates signaling by the T cell antigen receptor (TCR) and by the mast cell high affinity receptor for IgE (FcΞ΅RI) |
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Download this section (compressed HTML)Q: Should GRAP2 carry collagen-activated signaling pathway (GPVI) and CLEC-2 signaling terms, given that Gads is needed only for responses to weak GPVI/CLEC-2 stimulation and is largely redundant with GRB2 in platelets (PMID:18826392, PMID:31948362)?
Q: Several reports (PMID:9857184, PMID:11034310) place Mona/GADS downstream of the M-CSF receptor in monocyte/macrophage development; is there loss-of-function evidence that would justify a myeloid receptor signaling term in addition to the antigen-receptor pathways?
Q: GOA has no T cell receptor signaling pathway annotation for GRAP2 despite the knockout and Jurkat-knockout data; are the NEW TCR signaling, FcERI signaling and TCR signalosome rows acceptable to GO curators?
Q: Early work reported that phospho-LAT associates with Grb2 but not GrpL (PMID:10209041), whereas later studies consistently show SH2-dependent GADS binding to LAT pY171/pY191; is the early negative result explained by lysis conditions or antibody choice?
Q: GADS binds FLT3 pY955/pY969 and potentiates FLT3-ITD mitogenic signaling in engineered cells and xenografts (PMID:26895103); is there evidence from primary myeloid cells or AML samples that would justify a FLT3 signaling annotation, or is this restricted to overexpression systems?
Q: GADS-knockout Jurkat cells preserve early PLCG1 phosphorylation and calcium flux (PMID:37006259) whereas GADS-suppressed HuT78 cells lose calcium flux (PMID:25636200); which compensating adaptors (GRB2, CD6-associated complexes) explain the difference, and which model reflects primary human T cells?
Experiment: In GADS-knockout Jurkat or primary human T cells, reconstitute wild-type GADS, a dimerization-defective mutant, Y45F and T262A, and measure LAT-SLP-76 coupling, PLCG1 phosphorylation, calcium flux and CD69 induction across a titration of TCR stimulus strength.
Hypothesis: GADS dimerization and linker/N-SH3 phosphorylation set the TCR stimulus threshold by controlling cooperative binding of GADS to multiple LAT phosphotyrosines.
Type: genetic complementation with domain mutants
Experiment: Use proximity labelling (TurboID-GADS) in resting and TCR- or FcERI-stimulated cells to identify GADS partners other than SLP-76 and LAT, comparing wild-type and SH2 (R-to-K) or C-SH3 mutants.
Hypothesis: GADS has partners beyond SLP-76 and LAT (e.g. HPK1, CD6) that contribute to branch-specific signaling and to its distinct phenotype compared with SLP-76 or LAT loss.
Type: proximity proteomics
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