Glyceraldehyde-3-phosphate dehydrogenase 2 (Tdh2, YJR009C) is one of three paralogous NAD+-dependent phosphorylating glyceraldehyde-3-phosphate dehydrogenases (GAPDH, EC 1.2.1.12; TDH1, TDH2, TDH3) of Saccharomyces cerevisiae. These homotetrameric enzymes catalyse the reversible oxidative phosphorylation of D-glyceraldehyde 3-phosphate to 1,3-bisphosphoglycerate with reduction of NAD+ to NADH, the sixth step of glycolysis and an obligatory step of gluconeogenesis. Tdh2 contributes about 25-30% of cellular GAPDH activity in growing cells on both glucose and ethanol, forms catalytically active homotetramers with higher apparent Vmax than Tdh3, and overlaps functionally with Tdh3 as a major glycolytic isoform. It is predominantly cytosolic, with additional pools in the cell wall and, in high-throughput studies, the nucleus, mitochondria and lipid particles. GAPDH S-nitrosation has been linked to oxidative stress-induced cell death in yeast.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | IBA GO_REF:0000033 | ACCEPT | Summary: PAINT IBA from GAPDH node PTN000089475, seeded by the three yeast TDH paralogs among other experimentally characterised GAPDHs. TDH2 is an active NAD-dependent phosphorylating GAPDH, so the inherited activity is correct; TDH2 appearing among the donors reflects its own experimental grounding. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 The contribution of the TDH1, TDH2, and TDH3 gene products to the total glyceraldehyde-3-phosphate dehydrogenase activity in wild type cells is 10-15, 25-30, and 50-60%, respectively |
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | IDA PMID:3905788 Differential expression of the three yeast glyceraldehyde-3-... | ACCEPT | Summary: McAlister and Holland used tdh insertion mutants to partition total GAPDH activity among the three genes; TDH2 encodes a major (25-30%) isoform of cellular GAPDH activity and the authors conclude all three genes encode catalytically active enzyme. Supporting Evidence: PMID:3905788 The contribution of the TDH1, TDH2, and TDH3 gene products to the total glyceraldehyde-3-phosphate dehydrogenase activity in wild type cells is 10-15, 25-30, and 50-60%, respectively PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme PMID:3905788 The polypeptides encoded by the TDH2 or TDH3 structural genes form catalytically active homotetramers |
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | IEA GO_REF:0000120 | ACCEPT | Summary: EC 1.2.1.12 / RHEA:10300 mapping from the UniProt catalytic-activity annotation; correct for TDH2. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme |
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA (GLUCFERMEN-PWY, superpathway of glucose fermentation) assigning the reversible GAP + NAD+ + Pi <-> 1,3-bisphosphoglycerate step (EC 1.2.1.12) to TDH2; the activity is experimentally established for TDH2. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme |
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA (GLUCONEO-PWY-1, gluconeogenesis I) assigning the reversible GAP + NAD+ + Pi <-> 1,3-bisphosphoglycerate step (EC 1.2.1.12) to TDH2; the activity is experimentally established for TDH2. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme |
| GO:0004365 glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA (GLYCOLYSIS, glycolysis I) assigning the reversible GAP + NAD+ + Pi <-> 1,3-bisphosphoglycerate step (EC 1.2.1.12) to TDH2; the activity is experimentally established for TDH2. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme |
| GO:0005515 protein binding | IPI PMID:15699485 Analysis of polyubiquitin conjugates reveals that the Rpn10 ... | REMOVE | Summary: IPI with ubiquitin (UBI4, P0CG63) from Mayor et al., who purified His6-ubiquitin conjugates and identified candidate proteasome substrates by MS. Tdh2 recovery reflects covalent ubiquitylation (being a substrate), not a binding function; protein binding is uninformative. Reason: Protein binding is uninformative and here reflects covalent ubiquitylation (substrate status), not a binding activity; removal does not imply Tdh2 is not ubiquitylated. Supporting Evidence: PMID:15699485 Using this approach, we identified 127 proteins that are candidate substrates for the 26S proteasome. |
| GO:0005634 nucleus | HDA PMID:26928762 One library to make them all: streamlining the creation of y... | KEEP AS NON CORE | Summary: SWAT-GFP library imaging assigned a nuclear signal for Tdh2 in addition to cytosol. Nuclear pools of GAPDH are described in many eukaryotes, but no nuclear function is established for yeast Tdh2. Reason: HTP localization; secondary pool, non-core. Supporting Evidence: PMID:26928762 we only assigned localizations that could be easily discriminated by eye: ER, nuclear periphery, cytosol, cell periphery, vacuole lumen, vacuole membrane, mitochondria, nucleus, bud/bud neck and punctate |
| GO:0005737 cytoplasm | HDA PMID:11914276 Subcellular localization of the yeast proteome. | ACCEPT | Summary: Kumar et al. genome-wide immunolocalization of epitope-tagged proteins placed Tdh2 in the cytoplasm; consistent with its glycolytic role. Supporting Evidence: PMID:11914276 By high-throughput immunolocalization of tagged gene products, we have determined the subcellular localization of 2744 yeast proteins. |
| GO:0005737 cytoplasm | IEA GO_REF:0000044 | ACCEPT | Summary: UniProt subcellular-location mapping (SL-0086 Cytoplasm); correct for TDH2, which is detected in cytosolic extracts. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0005737 cytoplasm | ISS GO_REF:0000024 | ACCEPT | Summary: ISS from Tdh1 (P00360) cytoplasmic localization; Tdh2 is >88% identical and is directly detected in cytosolic extracts. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0005739 mitochondrion | IDA PMID:16962558 Enolase takes part in a macromolecular complex associated to... | KEEP AS NON CORE | Summary: Brandina et al. report that all glycolytic enzyme activities, including GAPDH, are partly associated with purified mitochondria and that glycolytic enzymes co-purify with an enolase-containing complex at the mitochondrial surface. This is a peripheral association of a small pool of TDH2; the abstract does not resolve individual TDH isoforms, but the curator had full text. Reason: Experimental, not removed; peripheral pool, not the site of core glycolytic activity. Supporting Evidence: PMID:16962558 we show that all glycolytic enzymes are associated with mitochondria in yeast PMID:16962558 The identified components included additional glycolytic enzymes |
| GO:0005739 mitochondrion | IEA GO_REF:0000117 | KEEP AS NON CORE | Summary: ARBA mapping to mitochondrion for GAPDH. Consistent with experimental detection of a peripheral (outer-surface) mitochondria-associated pool of GAPDH, but this is condition-dependent and not the site of core activity. Reason: Peripheral location propagated electronically; non-core. Supporting Evidence: PMID:16962558 we show that all glycolytic enzymes are associated with mitochondria in yeast |
| GO:0005811 lipid droplet | IDA PMID:10515935 Identification and characterization of major lipid particle ... | KEEP AS NON CORE | Summary: Athenstaedt et al. identified GAPDH by MS as a minor component in a gel band of highly purified lipid particles (less abundant than Yju3p). A minor co-purifying pool of an abundant cytosolic enzyme; no role at lipid droplets is described. Reason: Experimental but minor co-fractionation; non-core. Supporting Evidence: PMID:10515935 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Yju3p (the amount of Yju3p was greater than GAPDH) |
| GO:0005811 lipid droplet | IEA GO_REF:0000117 | KEEP AS NON CORE | Summary: ARBA rule propagating lipid droplet location to GAPDHs, derived from proteomic detection of GAPDH in lipid-particle fractions; consistent with the experimental row but a minor co-fractionating pool, not a characteristic location. Reason: Minor co-fractionation location; non-core. Supporting Evidence: PMID:10515935 glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Yju3p (the amount of Yju3p was greater than GAPDH) |
| GO:0005829 cytosol | HDA PMID:26928762 One library to make them all: streamlining the creation of y... | ACCEPT | Summary: SWAT-GFP library imaging (Meurer/Weill et al.) assigned TDH2 to the cytosol; consistent with its glycolytic role. Supporting Evidence: PMID:26928762 we only assigned localizations that could be easily discriminated by eye: ER, nuclear periphery, cytosol, cell periphery, vacuole lumen, vacuole membrane, mitochondria, nucleus, bud/bud neck and punctate |
| GO:0005829 cytosol | IBA GO_REF:0000033 | ACCEPT | Summary: PAINT IBA is_active_in cytosol for glycolytic GAPDHs; correct for TDH2. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0005829 cytosol | IDA PMID:11158358 The glyceraldehyde-3-phosphate dehydrogenase polypeptides en... | ACCEPT | Summary: Delgado et al. detected Tdh2 polypeptide by Western blot in cytosol of exponentially growing cells; cytosol is the site of GAPDH activity. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0005829 cytosol | IEA GO_REF:0000117 | ACCEPT | Summary: ARBA mapping to cytosol; correct for TDH2. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0005829 cytosol | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA is_active_in cytosol for the GAPDH step of superpathway of glucose fermentation; TDH2 is a soluble cytosolic glycolytic/gluconeogenic enzyme. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in |
| GO:0005829 cytosol | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA is_active_in cytosol for the GAPDH step of gluconeogenesis I; TDH2 is a soluble cytosolic glycolytic/gluconeogenic enzyme. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in |
| GO:0005829 cytosol | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA is_active_in cytosol for the GAPDH step of glycolysis I; TDH2 is a soluble cytosolic glycolytic/gluconeogenic enzyme. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in |
| GO:0005886 plasma membrane | HDA PMID:16622836 The plasma membrane proteome of Saccharomyces cerevisiae and... | KEEP AS NON CORE | Summary: Delom et al. identified ~90 proteins in a stripped plasma-membrane fraction by IEC/LDS-PAGE MS, including TDH2. Abundant glycolytic enzymes are common contaminants of membrane fractions, although yeast GAPDH is also found at the cell surface/cell wall (PMID:11158358). Reason: HTP proteomic detection; plausible peripheral pool, non-core. Supporting Evidence: PMID:16622836 Proteins from a stripped plasma membrane fraction were solubilized with the neutral and non-denaturing detergent, the n-dodecyl beta-D-maltoside. PMID:11158358 By immunoelectron microscopy, the GAPDH protein was detected at the outer surface of the cell wall of wild-type cells, as well as in the cytoplasm |
| GO:0006006 glucose metabolic process | IEA GO_REF:0000002 | TOO GENERAL | Summary: InterPro IPR006424 (type I GAPDH) mapping to glucose metabolic process. Correct but very general; the specific processes glycolysis and gluconeogenesis are annotated separately. Reason: Correct, general parent-level process; redundant with glycolysis/gluconeogenesis. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in |
| GO:0006094 gluconeogenesis | IEA GO_REF:0000117 | ACCEPT | Summary: ARBA mapping to gluconeogenesis. GAPDH catalyses the reversible step used in both glycolysis and gluconeogenesis, and TDH2 is expressed on ethanol as well as glucose. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 The relative proportions of expression of each gene is the same in cells grown in the presence of glucose or ethanol as carbon source |
| GO:0006094 gluconeogenesis | IEP PMID:3905788 Differential expression of the three yeast glyceraldehyde-3-... | ACCEPT | Summary: McAlister and Holland measured TDH2 expression/activity in ethanol-grown (gluconeogenic) as well as glucose-grown cells. IEP is weak evidence for participation, but the reversible GAPDH step is an obligatory part of gluconeogenesis, so the process assignment is correct. Supporting Evidence: PMID:3905788 The relative proportions of expression of each gene is the same in cells grown in the presence of glucose or ethanol as carbon source |
| GO:0006094 gluconeogenesis | RCA GO_REF:0000123 | ACCEPT | Summary: YeastPathways RCA (gluconeogenesis I) for TDH2; the GAPDH step operates in the gluconeogenic direction during growth on non-fermentable carbon sources. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 The relative proportions of expression of each gene is the same in cells grown in the presence of glucose or ethanol as carbon source |
| GO:0006096 glycolysis | IBA GO_REF:0000033 | ACCEPT | Summary: PAINT IBA for glycolysis from the GAPDH clade; TDH2 contributes a major (25-30%) isoform of the glycolytic GAPDH activity. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt glyceraldehyde 3-phosphate: step 1/5. {ECO:0000269|PubMed:3905788}. PMID:3905788 The contribution of the TDH1, TDH2, and TDH3 gene products to the total glyceraldehyde-3-phosphate dehydrogenase activity in wild type cells is 10-15, 25-30, and 50-60%, respectively |
| GO:0006096 glycolysis | IEA GO_REF:0000120 | ACCEPT | Summary: UniPathway UPA00109 / ARBA mapping to glycolysis; correct for TDH2. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt glyceraldehyde 3-phosphate: step 1/5. {ECO:0000269|PubMed:3905788}. |
| GO:0006096 glycolysis | IEP PMID:3905788 Differential expression of the three yeast glyceraldehyde-3-... | ACCEPT | Summary: McAlister and Holland: total GAPDH activity is 2-fold higher in glucose-grown cells, and TDH2 supplies a major (25-30%) isoform of it. IEP is indirect, but the glycolytic role of TDH2 is well established. Supporting Evidence: PMID:3905788 The contribution of the TDH1, TDH2, and TDH3 gene products to the total glyceraldehyde-3-phosphate dehydrogenase activity in wild type cells is 10-15, 25-30, and 50-60%, respectively PMID:3905788 The relative proportions of expression of each gene is the same in cells grown in the presence of glucose or ethanol as carbon source |
| GO:0006915 apoptotic process | IMP PMID:17726063 NO-mediated apoptosis in yeast. | KEEP AS NON CORE | Summary: Almeida et al. linked H2O2-induced, NO-dependent yeast apoptosis to GAPDH S-nitrosation and proteolysis; the IMP presumably reflects altered death/survival of a tdh mutant (full text not available here). GAPDH appears as a target/mediator of the death programme rather than a dedicated apoptotic effector, and the phenotype may be partly metabolic. Reason: Experimental (curator saw full text), not removed; stress/death role is peripheral to the core glycolytic function. Supporting Evidence: PMID:17726063 yeast GAPDH is a target of extensive proteolysis upon H2O2-induced apoptosis and undergoes S-nitrosation PMID:17726063 NO signalling and GAPDH S-nitrosation are linked with H2O2-induced apoptotic cell death |
| GO:0009277 fungal-type cell wall | IDA PMID:11158358 The glyceraldehyde-3-phosphate dehydrogenase polypeptides en... | KEEP AS NON CORE | Summary: Delgado et al. detected catalytically active GAPDH at the outer surface of the cell wall by immunofluorescence, immuno-EM and Western blotting of cell-wall extracts; TDH2 polypeptide was specifically detected in cell-wall extracts. Reason: Experimentally supported secondary (moonlighting) location; function there unknown. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells PMID:11158358 By immunoelectron microscopy, the GAPDH protein was detected at the outer surface of the cell wall of wild-type cells, as well as in the cytoplasm |
| GO:0009277 fungal-type cell wall | IEA GO_REF:0000117 | KEEP AS NON CORE | Summary: ARBA mapping to fungal-type cell wall; consistent with direct experimental detection of TDH2 in cell-wall extracts. Reason: Secondary location, non-core. Supporting Evidence: PMID:11158358 Western immunoblot analysis of cell wall extracts and cytosol showed that Tdh2 and Tdh3 polypeptides are present in the cell wall, as well as in the cytosol, of exponentially growing cells |
| GO:0016620 oxidoreductase activity, acting on the aldehyde or oxo group of donors, NAD or NADP as acceptor | IEA GO_REF:0000002 | MODIFY | Summary: InterPro GAPDH signatures mapped to the generic parent oxidoreductase activity acting on aldehyde/oxo donors with NAD(P) acceptor. Correct but uninformative; the specific child term GAPDH (NAD+) (phosphorylating) activity is supported. Reason: Too general; replace with the specific activity. Proposed replacements: glyceraldehyde-3-phosphate dehydrogenase (NAD+) (phosphorylating) activity Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in PMID:3905788 These data confirm that the three yeast glyceraldehyde-3-phosphate dehydrogenase genes encode catalytically active enzyme |
| GO:0019658 bifid shunt | RCA GO_REF:0000123 | REMOVE | Summary: YeastPathways RCA from GLUCFERMEN-PWY mapped to GO bifid shunt, the Bifidobacterium phosphoketolase pathway converting glucose to lactate and acetate. S. cerevisiae has no bifid shunt; TDH2 acts in glycolysis/alcoholic fermentation. Reason: Mis-mapped pathway: bifid shunt does not occur in S. cerevisiae. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt glyceraldehyde 3-phosphate: step 1/5. {ECO:0000269|PubMed:3905788}. |
| GO:0050661 NADP binding | IEA GO_REF:0000002 | MARK AS OVER ANNOTATED | Summary: InterPro IPR006424 (type I GAPDH family, which includes NADP-dependent plant/cyanobacterial GapB/GapC) mapped to NADP binding. Yeast TDH2 is the NAD+-specific phosphorylating GAPDH (EC 1.2.1.12); the only NADP(H) interaction recorded is a slow NAD(P)H-hydratase side activity, so NADP binding is not a meaningful functional description. Reason: Family-level mapping that over-states cofactor specificity of an NAD-dependent GAPDH. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt FUNCTION: As a side activity, catalyzes the hydration of the file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in |
| GO:0051287 NAD binding | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: InterPro GAPDH NAD(P)-binding domain mapped to NAD binding; correct, NAD+ is the cosubstrate of TDH2. Part of the catalytic activity rather than a separate function. Reason: Cosubstrate binding; non-core. Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) involved in file:yeast/TDH2/TDH2-uniprot.txt KM=0.37 uM for NAD(+) |
| GO:0061620 obsolete glycolytic process through glucose-6-phosphate | RCA GO_REF:0000123 | MODIFY | Summary: YeastPathways RCA (GLYCOLYSIS) to glycolytic process through glucose-6-phosphate, now obsolete and replaced by glycolysis (GO:0006096). TDH2 does act in glycolysis. Reason: Obsolete term; use replaced_by GO:0006096. Proposed replacements: glycolysis Supporting Evidence: file:yeast/TDH2/TDH2-uniprot.txt glyceraldehyde 3-phosphate: step 1/5. {ECO:0000269|PubMed:3905788}. |
| GO:0072593 reactive oxygen species metabolic process | IMP PMID:17726063 NO-mediated apoptosis in yeast. | KEEP AS NON CORE | Summary: Almeida et al.: blocking NO synthesis decreases GAPDH S-nitrosation and intracellular ROS accumulation; the curator's IMP links TDH2 loss to altered ROS levels. Likely an indirect consequence of redox/metabolic perturbation rather than a dedicated ROS-metabolising activity. Reason: Indirect/pleiotropic effect; non-core. Supporting Evidence: PMID:17726063 Blockage of NO synthesis with Nomega-nitro-L-arginine methyl ester leads to a decrease of GAPDH S-nitrosation and of intracellular reactive oxygen species (ROS) accumulation, increasing survival PMID:17726063 NO signalling and GAPDH S-nitrosation are linked with H2O2-induced apoptotic cell death |
| GO:1904408 melatonin binding | IDA PMID:31708896 Glycolytic Proteins Interact With Intracellular Melatonin in... | MARK AS OVER ANNOTATED | Summary: Morcillo-Parra et al. immunopurified melatonin-associated proteins from fermenting cells with anti-melatonin IgG beads and identified Tdh1/2/3 by MS in one gel band. This is an affinity co-purification without a defined binding site, stoichiometry or physiological effect on TDH2. Reason: Affinity co-purification of abundant GAPDH paralogs is weak support for a specific melatonin-binding activity; the experimental report is retained but likely overstates a physiologic molecular function. Supporting Evidence: PMID:31708896 Surprisingly, most of the proteins identified participate in the glycolytic pathway: pyruvate kinase 1 (Pyk2p; band a), enolase (Eno1p, Eno2p; band c), fructose biphosphate aldolase (Fba1p; band d), and glyceraldehyde-3-phosphate dehydrogenase (Tdh1p, Tdh2p, Tdh3p; band f). PMID:31708896 We subsequently performed immunopurification melatonin-IgG-Dynabeads and identified a set of proteins that interact with melatonin inside the cell. |
| GO:1904408 melatonin binding | IEA GO_REF:0000117 | MARK AS OVER ANNOTATED | Summary: ARBA rule propagating melatonin binding to GAPDHs, derived from a single affinity pull-down study in yeast that did identify this protein; physiological significance is unknown. Reason: ARBA propagates the same weak affinity-capture result as a molecular-function assertion; the interaction lacks a binding constant, site, or demonstrated physiologic effect. Supporting Evidence: PMID:31708896 We subsequently performed immunopurification melatonin-IgG-Dynabeads and identified a set of proteins that interact with melatonin inside the cell. |
| GO:1990841 promoter-specific chromatin binding | IDA PMID:27184763 HyCCAPP as a tool to characterize promoter DNA-protein inter... | MARK AS OVER ANNOTATED | Summary: HyCCAPP (hybridization capture of chromatin-associated proteins) identified Tdh2 at the ENO2 promoter, and ChIP showed Tdh2 enrichment at the GAL1 locus with no difference between glucose and galactose. GAPDH has no known DNA-binding domain in yeast and is one of the most abundant cellular proteins; condition-independent crosslinking to chromatin does not establish promoter-specific binding activity. Reason: Crosslinking-based association without specificity or functional consequence; likely reflects protein abundance rather than a promoter-binding activity. Supporting Evidence: PMID:27184763 Both Tdh2 and Rpa190 showed enrichment compared to the negative control, but no significant difference between the two growth conditions PMID:27184763 Both Pab1 and Tdh2, targeted as part of the ENO2 HyCCAPP validation, were analyzed, and showed enrichment at the GAL1 locus. |
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Experiment: ChIP of Tdh2 at GAL1/ENO2 promoters normalised to a highly expressed non-DNA-binding cytosolic control protein, with catalytic-site and surface mutants, to test whether chromatin association is specific.
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), isozyme 2; involved in glycolysis and gluconeogenesis; tetramer that catalyzes reaction of glyceraldehyde-3-phosphate to 1,3 bis-phosphoglycerate; detected in cytoplasm and cell wall; protein abundance increases in response to DNA replication stress; GAPDH-derived antimicrobial peptides are active against a wide variety of wine-related yeasts and bateria; TDH2 has a paralog, TDH3, that arose from the whole genome duplication
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