ASNS

Existing Annotations Review

GO Term	Evidence	Action	Reason
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IBA GO_REF:0000033	ACCEPT	Summary: ASNS catalyzes glutamine-dependent asparagine synthesis. Reason: The inherited catalytic assertion agrees with direct human recombinant-enzyme and mutagenesis studies. The target appearing among the descendant experimental sources is expected for PAINT and is not circular evidence. No target-specific loss of this activity is evident. Human primary abstracts and targeted 2024 enzymology read; the actual ancestral-node placement/MSA was not independently reconstructed. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro. PMID:2573597 Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IDA PMID:16023613 Efficient expression, purification, and characterization of ...	ACCEPT	Summary: Purified recombinant human ASNS retains glutamine-dependent synthetase activity. Reason: The cited study reports active C-terminally tagged human enzyme produced in a baculovirus-based system, including correct processing. This supports the defining catalytic activity together with direct human reaction evidence; the tag does not warrant changing the original gene-level assertion. Complete cited abstract; original detailed purification and assay Methods not read. Supporting Evidence: PMID:16023613 We now report an efficient protocol for preparing multi-milligram quantities of C-terminally tagged, wild type human AS in a baculovirus-based expression system. The recombinant enzyme is correctly processed and exhibits high catalytic activity. PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IDA PMID:2564390 Expression of human asparagine synthetase in Escherichia col...	ACCEPT	Summary: Human ASNS expressed in bacteria produces active asparagine synthetase. Reason: The cited human recombinant study reports both glutamine-dependent and ammonia-dependent asparagine-synthesis activity. Retain the specific glutamine-hydrolyzing MF as the principal biochemical function. Complete cited abstract; no claim of reading the original detailed kinetic tables. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IDA PMID:2569668 Organization and expression of the cell cycle gene, ts11, th...	ACCEPT	Summary: Human ts11 encodes the asparagine-synthesizing enzyme ASNS. Reason: The cited study identifies the human gene through BHK ts11 complementation and asparagine bypass. The exact original IDA enzyme-assay passage is not available in the abstract, but the assigned catalytic function is independently established by human biochemical studies; retain the curator assertion without relabeling the complementation experiment as a purified assay. Complete cited abstract; original IDA assay details deferred to the curator, independently corroborated by PMID:2564390 / PMID:2573597 / PMID:16023613. Supporting Evidence: PMID:2569668 The human ts11 gene was isolated on the basis of its ability to complement the mutation of the BHK cell cycle ts11 mutant, which is blocked in G1 at the nonpermissive temperature. This gene has now been identified as the structural gene for asparagine synthetase (AS) on the bases of sequence homology and the ability of exogenous asparagine to bypass the ts11 block. PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IDA PMID:2573597 The N-terminal cysteine of human asparagine synthetase is es...	ACCEPT	Summary: The N-terminal catalytic cysteine is required for glutamine-dependent asparagine synthesis. Reason: Replacing the N-terminal cysteine with alanine in human ASNS expressed in yeast abolishes glutamine-dependent activity while leaving ammonia-dependent activity intact. This supports the coupled catalytic function and the specialized N-terminal glutamine-amidotransfer step. Complete cited abstract, which explicitly describes the human construct, yeast expression and in-vitro discrimination. Supporting Evidence: PMID:2573597 Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IDA PMID:2886907 Isolation of human cDNAs for asparagine synthetase and expre...	ACCEPT	Summary: Human ASNS cDNA restores asparagine synthesis in deficient recipient cells. Reason: The cited human fibroblast cDNA is expressed in Jensen rat sarcoma cells and rescues asparagine auxotrophy. Retain the established human enzyme function; recipient-species context and the unread detailed enzyme-assay methods are explicit. Complete cited abstract; human cDNA and rat recipient distinguished. Supporting Evidence: PMID:2886907 The cDNAs containing the entire protein-coding sequence expressed asparagine synthetase activity and were capable of conferring asparagine prototrophy on the Jensen rat sarcoma cells.
GO:0004066 asparagine synthase (glutamine-hydrolyzing) activity	IEA GO_REF:0000120	ACCEPT	Summary: Sequence and reaction mappings identify the principal ASNS catalytic activity. Reason: The Rhea/EC/domain and orthology mappings converge on the directly established human glutamine-dependent reaction. The original electronic evidence and all WITH/FROM values are retained; support is not based only on a family label. Exact mapped reaction and human biochemical evidence read; individual automated rule internals not reconstructed. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.
GO:0004071 aspartate-ammonia ligase activity	IEA GO_REF:0000116	KEEP AS NON CORE	Summary: Human ASNS can synthesize asparagine using free ammonia in vitro. Reason: GO:0004071 specifies ammonia-fed synthesis, distinct from the glutamine-fed GO:0004066 reaction. Human recombinant studies and the 2024 wild-type kinetic table support this activity. Retain it as a measured alternative substrate mode, without asserting that free ammonia is the usual physiological nitrogen donor or creating a second core. Current GO definition/parent, original 1989 abstract and targeted 2024 Table 1/kinetic Methods; actual normal XML cache now read, including expression/purification Methods and the kinetic assay. The 2024 assay measures PPi formation, with asparagine stoichiometry inferred by analogy to the bacterial homolog; free-ammonia activity is an experimental substrate mode. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro. PMID:2573597 Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected. PMID:39627226 When ammonia was used as the nitrogen source, varying the ammonia concentration gave steady-state parameters for the two variants that were essentially unchanged compared to those of WT ASNS (Table 1 and Supplementary Fig. 32). PMID:39627226 The rate of inorganic pyrophosphate (PPi) production catalyzed by recombinant, WT human ASNS and two ammonia tunnel variants was determined in a continuous assay format using the EnzChek™ Pyrophosphate Assay Kit (Molecular Probes, Eugene, OR).
GO:0004359 glutaminase activity	IEA GO_REF:0000120	ACCEPT	Summary: The N-terminal ASNS catalytic domain hydrolyzes glutamine to supply nitrogen for asparagine synthesis. Reason: ASNS hydrolyzes glutamine at its N-terminal catalytic site to supply ammonia to the C-terminal asparagine-synthesis reaction. This is an integral catalytic step of its defining glutamine-dependent biosynthesis function. Human N-terminal cysteine mutagenesis selectively eliminates glutamine-dependent synthesis while preserving ammonia-fed synthesis, and the human structure identifies the two catalytic domains and connecting tunnel. ACCEPT retains this core half-reaction; the single synthesized core represents the complete coupled reaction GO:0004066. RHEA:15889 is curator-inferred in the shipped UniProt record, and no independently measured autonomous glutaminase assay or bulk glutamine-catabolic role is claimed. Supporting Evidence: PMID:2573597 Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected. file:human/ASNS/ASNS-uniprot.txt CC Reaction=L-glutamine + H2O = L-glutamate + NH4(+); CC Xref=Rhea:RHEA:15889, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938, CC ChEBI:CHEBI:29985, ChEBI:CHEBI:58359; EC=3.5.1.2; PMID:39627226 a Schematic representation showing the N-terminal glutaminase (forest green), dimerization (magenta) and the C-terminal synthetase (cornflower blue) domains.
GO:0005515 protein binding	IPI PMID:25416956 A proteome-scale map of the human interactome network.	REMOVE	Summary: Remove uninformative bare protein binding while preserving the experimentally reported ASNS–WDR27 interaction. Reason: The source-specific ASNS-bait/WDR27-prey pair was established in the prior BioGRID/PMID:25416956 audit and remains valid interaction evidence. The inspected pair supports an association but no more informative ASNS molecular activity, so remove the uninformative bare-binding term without asserting that the interaction is false. The three TRIM69 rows are held under the explicit source-adjudication exception: their distinct source/variant/isoform records remain unread, preventing a source-specific decision about any justified refinement. This is not a general requirement to disprove binding before removing GO:0005515, and no ligase-binding term is inferred from a partner name. Supporting Evidence: file:human/ASNS/ASNS-notes.md The exact human ASNS-bait/WDR27-prey two-hybrid record was independently read in [BioGRID interaction 1037059](https://thebiogrid.org/interaction/1037059/asns-wdr27.html). Its publication link resolves to the Rolland 2014 record and explicitly PMID:25416956. This curated source-specific pair supports non-core retention, with the original supplementary constructs/raw reporter data unread.
GO:0005515 protein binding	IPI PMID:25416956 A proteome-scale map of the human interactome network.	UNDECIDED	Summary: The 2014 screen assigns an ASNS–TRIM69 interaction. Reason: The shipped source row and UniProt aggregation identify TRIM69 (Q86WT6), but the inspected 2014 article extraction and currently exposed portal did not resolve this exact source-specific pair/construct record. Retain the positive assertion pending that scope check; the missing table is not evidence that the pair is false. Complete paper abstract and available abbreviated HTML body; exact supplementary pair unexposed. No ligase-binding refinement is inferred from the partner name alone. The bare-binding term is uninformative regardless of whether the pair is real. UNDECIDED applies here under the local source-access exception because the exact source/construct assertion has not been adjudicated, including whether it warrants a specific refinement; it is not a claim that missing access disproves binding or that interaction confidence determines the usefulness of GO:0005515. The independently resolved WDR27 pair has no demonstrated more specific function and is removed under the default policy.
GO:0005515 protein binding	IPI PMID:31515488 Extensive disruption of protein interactions by genetic vari...	UNDECIDED	Summary: The 2019 variant screen assigns an ASNS–TRIM69 interaction. Reason: This study measures interaction effects of human sequence variants. The available main text establishes the screen and validation design, but the exact ASNS–TRIM69 reference-versus-variant pair and construct result remain unread. The aggregate UniProt interaction does not establish which result belongs to this paper. Complete abstract and targeted Results describing Y2H/orthogonal PCA validation; target supplementary row not read. The bare-binding term is uninformative regardless of whether the pair is real. UNDECIDED applies here under the local source-access exception because the exact source/construct assertion has not been adjudicated, including whether it warrants a specific refinement; it is not a claim that missing access disproves binding or that interaction confidence determines the usefulness of GO:0005515. The independently resolved WDR27 pair has no demonstrated more specific function and is removed under the default policy.
GO:0005515 protein binding	IPI PMID:32296183 A reference map of the human binary protein interactome.	UNDECIDED	Summary: HuRI assigns ASNS binding to the specified TRIM69 isoform. Reason: The original WITH/FROM identifier Q86WT6-2 is preserved. The available HuRI article and portal response did not expose the exact ASNS–TRIM69-2 dataset/construct entry; the isoform-specific source claim therefore remains unresolved without implying an absent interaction. Complete paper abstract and screen overview; primary portal TRIM69 page exposed an empty interaction table during this read. Exact target/isoform table not verified. The bare-binding term is uninformative regardless of whether the pair is real. UNDECIDED applies here under the local source-access exception because the exact source/construct assertion has not been adjudicated, including whether it warrants a specific refinement; it is not a claim that missing access disproves binding or that interaction confidence determines the usefulness of GO:0005515. The independently resolved WDR27 pair has no demonstrated more specific function and is removed under the default policy.
GO:0005829 cytosol	IBA GO_REF:0000033	ACCEPT	Summary: ASNS carries out its amino-acid biosynthetic reaction in the cytosol. Reason: The PAINT location/activity inference agrees with current human immunofluorescence and the explicit cytosolic ASNS catalyst in Reactome. The human target among descendant sources is appropriate; no donor-count or circularity objection is made. Current HPA gene table and normal Reactome summary read; precise ancestral-node placement not independently reconstructed. Supporting Evidence: Reactome:R-HSA-70599 Cytosolic asparagine synthase (ASNS) catalyzes the reaction of aspartate, glutamine, and ATP to form asparagine, glutamate, AMP, and pyrophosphate. file:human/ASNS/ASNS-notes.md The current [HPA ASNS subcellular page](https://www.proteinatlas.org/ENSG00000070669-ASNS/subcellular) reports Cytosol, Supported. Displayed antibody/cell-line records include HPA004924 in A431/U251MG/U2OS and HPA064737 in MCF7/SK-MEL-30/U2OS. The historical images were not independently rescored.
GO:0005829 cytosol	IDA GO_REF:0000052	ACCEPT	Summary: Human immunofluorescence localizes ASNS to the cytosol. Reason: The current HPA ASNS subcellular page reports supported cytosolic localization with two antibodies and several cell lines. This corroborates the original HPA IDA location while preserving its located_in relation; cytosolic localization is not asserted to be exclusive. Current HPA gene table and displayed antibody/cell-line localization entries read; original historical images not independently rescored. Supporting Evidence: file:human/ASNS/ASNS-notes.md The current [HPA ASNS subcellular page](https://www.proteinatlas.org/ENSG00000070669-ASNS/subcellular) reports Cytosol, Supported. Displayed antibody/cell-line records include HPA004924 in A431/U251MG/U2OS and HPA064737 in MCF7/SK-MEL-30/U2OS. The historical images were not independently rescored.
GO:0005829 cytosol	IEA GO_REF:0000107	ACCEPT	Summary: The mouse-to-human location transfer agrees with observed human cytosolic ASNS. Reason: The existing orthology-derived cytosolic activity annotation is concordant with current human localization and the cytosolic asparagine-synthesis reaction. Preserve the electronic source, donor and qualifier rather than replacing them with the independent human observations. Current human HPA and Reactome corroboration; original mouse assay and Ensembl transfer rule not independently rederived. Supporting Evidence: Reactome:R-HSA-70599 Cytosolic asparagine synthase (ASNS) catalyzes the reaction of aspartate, glutamine, and ATP to form asparagine, glutamate, AMP, and pyrophosphate. file:human/ASNS/ASNS-notes.md The current [HPA ASNS subcellular page](https://www.proteinatlas.org/ENSG00000070669-ASNS/subcellular) reports Cytosol, Supported. Displayed antibody/cell-line records include HPA004924 in A431/U251MG/U2OS and HPA064737 in MCF7/SK-MEL-30/U2OS. The historical images were not independently rescored.
GO:0005829 cytosol	TAS Reactome:R-HSA-70599	ACCEPT	Summary: Reactome places the ASNS-catalyzed synthesis reaction in the cytosol. Reason: The actual R-HSA-70599 summary explicitly identifies cytosolic ASNS catalysis of the glutamine/aspartate/ATP reaction. This is a target-specific catalytic location assertion, not a generic compartment inferred from pathway membership. Complete normal event summary and current official event page; original 1989 detailed fractionation Methods not claimed. Supporting Evidence: Reactome:R-HSA-70599 Cytosolic asparagine synthase (ASNS) catalyzes the reaction of aspartate, glutamine, and ATP to form asparagine, glutamate, AMP, and pyrophosphate.
GO:0042149 cellular response to glucose starvation	IDA PMID:10085239 Transcriptional regulation of the human asparagine synthetas...	KEEP AS NON CORE	Summary: ASNS expression increases during glucose deprivation in human cultured cells. Reason: The cited study measures ASNS mRNA induction in glucose-deprived human HepG2 and MOLT4 cells and promoter-dependent transcriptional induction. This fits the current cellular-response term, which includes gene-expression changes. Retain the existing contextual response without identifying ASNS as the upstream glucose sensor. Complete cited abstract; its repeated abstract under Full Text is not a full body. Current GO definition read. Supporting Evidence: PMID:10085239 Incubation of human HepG2 hepatoma cells in glucose-free medium resulted in an increased AS mRNA content, reaching a maximum of about 14-fold over control cells after approx. 12 h.
GO:0043066 negative regulation of apoptotic process	IMP PMID:17409444 Enhanced expression of asparagine synthetase under glucose-d...	KEEP AS NON CORE	Summary: ASNS protects pancreatic cancer cells against apoptosis under particular stress conditions. Reason: Own-study ASNS knockdown and overexpression alter apoptosis in MiaPaCa2 cells under glucose deprivation and cisplatin treatment. This supports a contextual anti-apoptotic role, rather than a universal survival function or direct assignment of a JNK enzymatic activity to ASNS. Complete cached abstract plus original author-upload apoptosis Methods, human ASNS transfection/siRNA Methods and targeted Results/Fig 2–3 passages. Cache remains abstract-only; no complete supplement read. Supporting Evidence: PMID:17409444 ASNS protects pancreatic cancer cells from apoptosis induced by glucose deprivation itself.
GO:0045931 positive regulation of mitotic cell cycle	IDA PMID:2569668 Organization and expression of the cell cycle gene, ts11, th...	KEEP AS NON CORE	Summary: Human ASNS supports progression through the cell cycle in the ts11 complementation system. Reason: Human ts11/ASNS complements the BHK G1 block, and exogenous asparagine bypasses that block. Retain the existing positive cell-cycle assertion as a contextual consequence of asparagine supply; do not infer a dedicated mitotic checkpoint or spindle mechanism from this paper. The precise original IDA assay and all cell-cycle experiments were not independently read. Complete own-study abstract with human gene, hamster recipient and human/mouse/hamster expression contexts; detailed full text unavailable. Supporting Evidence: PMID:2569668 The human ts11 gene was isolated on the basis of its ability to complement the mutation of the BHK cell cycle ts11 mutant, which is blocked in G1 at the nonpermissive temperature. This gene has now been identified as the structural gene for asparagine synthetase (AS) on the bases of sequence homology and the ability of exogenous asparagine to bypass the ts11 block.
GO:0070981 L-asparagine biosynthetic process	IBA GO_REF:0000033	ACCEPT	Summary: ASNS is an inherited catalyst of L-asparagine biosynthesis. Reason: The PAINT biological-process assertion matches the direct work of the enzyme: it catalyzes formation of asparagine rather than being merely a substrate or passive requirement. Human experimental evidence corroborates the conserved chemistry, with no proposed new process or donor-count objection. Exact existing IBA source set preserved; human enzymology and current BP definition read, ancestral placement not independently reconstructed. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro. Reactome:R-HSA-70599 Cytosolic asparagine synthase (ASNS) catalyzes the reaction of aspartate, glutamine, and ATP to form asparagine, glutamate, AMP, and pyrophosphate.
GO:0070981 L-asparagine biosynthetic process	IC PMID:2564390 Expression of human asparagine synthetase in Escherichia col...	ACCEPT	Summary: The directly demonstrated ASNS reaction produces L-asparagine. Reason: The IC assertion from GO:0004066 is justified because the human enzyme performs the asparagine-forming chemical step. Preserve the IC evidence and source rather than manufacturing a new experiment. Complete human recombinant-enzyme abstract and exact current MF/BP reactions. Supporting Evidence: PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.
GO:0070981 L-asparagine biosynthetic process	IDA PMID:2573597 The N-terminal cysteine of human asparagine synthetase is es...	ACCEPT	Summary: Human ASNS catalytic mutagenesis establishes its contribution to asparagine production. Reason: The own-source in-vitro human Cys-mutant experiment directly distinguishes the glutamine-dependent route to asparagine. Retain the existing acts_upstream_of_or_within qualifier exactly; no new qualifier or organism claim is imposed. Complete cited abstract; original source object including broader qualifier unchanged. Supporting Evidence: PMID:2573597 Site-specific mutagenesis was used to replace the N-terminal cysteine in human asparagine synthetase by an alanine. The mutant enzyme was expressed in the yeast Saccharomyces cerevisiae, and the asparagine synthetase activity was analyzed in vitro. The mutation resulted in the loss of the glutamine-dependent asparagine synthetase activity, while the ammonia-dependent activity remained unaffected.
GO:0070981 L-asparagine biosynthetic process	IEA GO_REF:0000120	ACCEPT	Summary: Electronic mappings correctly place ASNS in L-asparagine biosynthesis. Reason: The orthology, domain and UniPathway assignment is supported by direct human synthesis chemistry and the ASNS-catalyzed Reactome reaction. The enzyme performs the step itself; no additional broad amino-acid process term is needed. Exact existing mappings plus human primary abstracts and normal Reactome summary; individual automated rule provenance not rederived. Supporting Evidence: Reactome:R-HSA-70599 Cytosolic asparagine synthase (ASNS) catalyzes the reaction of aspartate, glutamine, and ATP to form asparagine, glutamate, AMP, and pyrophosphate. PMID:2564390 The recombinant enzyme was shown to have both the ammonia- and glutamine-dependent asparagine synthetase activity in vitro.

Gene Description

Existing Annotations Review

Core Functions

Couples N-terminal glutamine hydrolysis and intramolecular nitrogen transfer to ATP-dependent L-asparagine synthesis in the cytosol. The two-domain human enzyme functions as a homodimer.

References

Suggested Questions for Experts

📚 Additional Documentation

Notes

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