| Function category | Molecular mechanism (reaction/interaction/PTM) | Key residues/PTMs | Subcellular localization/compartment | Biological consequence | Key recent evidence (2023-2024) with DOI URL and date | Citation ID(s) |
|---|---|---|---|---|---|---|
| Canonical glycolysis | NAD+-dependent oxidation/phosphorylation of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate; active homotetramer | Catalytic cysteine reported as C150/C152; active-site region 150-157; NAD+-binding domain residues 1-150 | Predominantly cytosol | Core glycolytic ATP-generating pathway function | Lin 2024, *Molecules*, 2024-04, DOI: https://doi.org/10.3390/molecules29071573; Lévénès 2024 thesis-style summary, 2024 | (pqac-00000003, pqac-00000000) |
| Moonlighting | S-nitrosylation/oxidation at catalytic cysteine enables GAPDH-Siah1 complex formation and nuclear translocation | Cys150; Lys225 required for Siah1 binding (K225A abolishes binding); Lys160 implicated downstream | Cytosol to nucleus | Initiates N-GAPDH cascade linked to apoptosis/stress signaling; in microglia mediates stress-induced cognitive inflexibility | Ramos 2024, *Molecular Psychiatry*, 2024-04, DOI: https://doi.org/10.1038/s41380-024-02553-1; RR blocker used at 1 nM in BV2 cells and 0.25 mg/kg/day i.p. for 5 days in vivo | (pqac-00000005) |
| Moonlighting | Nuclear translocation consumes only a minor fraction of total GAPDH pool | N-GAPDH estimated at 1-2% of total GAPDH | Nuclear microglia (context-specific recent evidence) | Regulatory signaling with negligible impact on bulk glycolysis, supporting separation of moonlighting from metabolic function | Ramos 2024, *Molecular Psychiatry*, 2024-04, DOI: https://doi.org/10.1038/s41380-024-02553-1 | (pqac-00000005) |
| Moonlighting | Nuclear GAPDH participates in DNA damage response/base excision repair via interaction with DNA polymerase beta; Src-mediated phosphorylation promotes nuclear entry under DNA damage | Tyr41 phosphorylation (Src-mediated) | Nucleus, DNA lesion sites | Enhances Pol beta polymerase activity and BER; may support tumor progression | Lin 2024, *Molecules*, 2024-04, DOI: https://doi.org/10.3390/molecules29071573 | (pqac-00000003, pqac-00000007) |
| Moonlighting | Trans-nitrosylase and stress-signaling hub; interacts with p300, PARP1, SIRT1, HDAC2, DNA-PK and others after PTM-driven relocalization | S-nitrosylation; sulfhydration; O-GlcNAcylation at Thr227; acetylation at Lys160; PARylation-related interactions | Nucleus and cytosol | Links redox stress to transcriptional regulation, autophagy, PARP1 activation, and cell death/survival decisions | Lévénès 2024 thesis-style summary, 2024 | (pqac-00000000, pqac-00000001, pqac-00000002) |
| Moonlighting | RNA-binding/transcript-regulatory functions | RNA interactions noted with AU-rich RNA, tRNA, telomerase RNA, and ccn2 mRNA cis-element | Cytosol and nucleus | Post-transcriptional regulation beyond glycolysis | Lévénès 2024 thesis-style summary, 2024 | (pqac-00000001, pqac-00000002) |
| Moonlighting | PTM-driven autophagy regulation | Sulfhydration; O-GlcNAcylation; interactions involving SIRT1 and Rheb/mTOR pathways | Cytosol to nucleus | Can promote autophagy and metabolic stress adaptation | Lévénès 2024 thesis-style summary, 2024 | (pqac-00000000, pqac-00000002) |
| Canonical glycolysis / PTM-regulated | Crotonylation decreases catalytic activity in human ESCs | Crotonylation at K194 and K219; p300 acts as crotonyltransferase in presence of crotonyl-CoA | Not explicitly localized in snippet | Reduced glycolysis accompanies endodermal differentiation/metabolic switch | Zhang 2023, *Stem Cell Research & Therapy*, 2023-04, DOI: https://doi.org/10.1186/s13287-023-03290-y; crotonate lowered GAPDH activity by ~50%; 3-BrPA lowered activity by >50% and increased GATA6/SOX17 >30-fold | (pqac-00000006) |
| Canonical glycolysis / PTM-regulated | HDAC9-mediated deacetylation during rotavirus infection | K219 deacetylation; modified peptide AVGK(Acetyl)VIPELNGK; K219 was the only modified residue identified in sample | Caco-2 cell context; compartment not specified in snippet | Promotes rotavirus replication and alters glycolysis | Song 2024, *Virus Genes*, 2024-09, DOI: https://doi.org/10.1007/s11262-024-02104-4; observed 50 h after infection; glycolysis/gluconeogenesis among 17 enriched glycometabolism pathways; implicated set accounted for 42.08% of proteins in the cell dataset | (pqac-00000013, pqac-00000015) |
| Expression/implementation note | Highly expressed housekeeping/reference gene candidate, but expression stability depends on tissue/condition; pseudogenes complicate interpretation | Mean TPM often >45 for GAPDH; recommended highly expressed RGs >20 TPM; pseudogene/multi-mapping concerns noted | Broad tissue expression | Useful but not universal normalization control; requires validation in specific experimental settings | Tung 2024, *Scientific Reports*, 2024-05, DOI: https://doi.org/10.1038/s41598-024-63269-4; Zhang 2023 cautions GAPDH expression is tightly regulated/variable; Lévénès 2024 cites pseudogene-expression concerns | (pqac-00000016, pqac-00000006, pqac-00000014) |
| Expression/implementation note | Abundant expression varies by tissue | Example TPM values reported for human GAPDH: lymphocytes 6385 TPM, pancreas 328 TPM | Ubiquitous across tissues | Reinforces strong baseline expression but also substantial tissue-dependent range | Lévénès 2024 thesis-style summary, 2024 | (pqac-00000000) |


*Table: This table summarizes supported canonical and moonlighting functions of human GAPDH (UniProt P04406), highlighting key PTMs, localizations, biological effects, and recent 2023-2024 evidence. It is useful as a compact annotation aid for distinguishing core glycolytic activity from stress-responsive regulatory roles and for noting practical assay caveats.*