Affinage

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase · UniProt P04406

Length
335 aa
Mass
36.1 kDa
Annotated
2026-06-10
100 papers in source corpus 39 papers cited in narrative 37 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 9/9 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

GAPDH is a tetrameric glycolytic enzyme whose active-site cysteine (Cys152 in human; Cys150/Cys152 in rodent and yeast orthologs) functions as a versatile redox switch that couples metabolic flux to a diverse repertoire of non-glycolytic, signaling functions (PMID:7573405, PMID:37024754). Quantitatively, GAPDH together with PGK1 operates near equilibrium and defines a rate-controlling node of glycolysis: flux is buffered until GAPDH activity falls below ~19% of normal, after which glycolysis declines proportionally (PMID:33545174). The catalytic cysteine is the convergence point for many covalent and oxidative modifications that inactivate the enzyme and re-route metabolism: reversible S-nitrosylation by NO/iNOS/nNOS (PMID:7573405, PMID:16574384, PMID:16633896), S-glutathionylation and reversible oxidation by H2O2 (PMID:34332079, PMID:37024754), H2S-dependent sulfhydration (PMID:33459133), succination by dimethyl/monomethyl fumarate (structurally shown to block NAD+ binding) (PMID:29599194, PMID:31387164), and alkylation at Cys22 by itaconate (PMID:31704924); the enzyme is additionally regulated by CARM1 methylation at Arg234 (PMID:30232003), δPKC phosphorylation that disrupts tetramerization (PMID:27129213), VRK2/FBXW10-dependent phosphorylation/ubiquitination at Ser151 (PMID:37450367), and TGM2-catalyzed serotonylation at Gln262 (PMID:38215751). These modifications loss of glycolytic activity divert carbon toward the oxidative pentose phosphate pathway and reductive metabolism under oxidative stress, a switch required for anchorage-independent and tumor growth (PMID:37024754). Inactivation of GAPDH underlies the anti-inflammatory actions of fumarate and itaconate by suppressing glycolysis in immune cells (PMID:29599194, PMID:31704924) and the selective vulnerability of glycolytic KRAS/BRAF-mutant tumors to oxidative GAPDH inactivation (PMID:26541605). Modification of the catalytic cysteine also drives nuclear translocation: S-nitrosylated GAPDH binds the E3 ligase Siah1 and is carried to the nucleus, where it is acetylated at Lys160 by p300/CBP, stimulates p300/CBP activity and p53-dependent apoptosis, and trans-modifies nuclear targets including SIRT1, HDAC2 and DNA-PK (PMID:16574384, PMID:16633896, PMID:18552833, PMID:20972425, PMID:24670206, PMID:34332079); this axis drives neuronal death and Aβ-induced tau acetylation in Alzheimer models, and is restrained by the cytosolic anchor GOSPEL (PMID:19607794, PMID:29559585). Beyond apoptosis, nuclear GAPDH supports base excision repair through DNA polymerase β following Src-mediated Tyr41 phosphorylation (PMID:32539222), partners with NAMPT to sustain nuclear NAD+ salvage (PMID:31988240), and binds the telomerase RNA TERC to inhibit telomerase (PMID:22847419). GAPDH additionally moonlights as a heme chaperone delivering heme to apo-sGCβ1, IDO1, TDO, and globins in complex with Hsp90 (PMID:32358060, PMID:34972240, PMID:35051612), and as a cell-surface receptor for transferrin/lactoferrin (iron homeostasis) and plasminogen (macrophage migration) (PMID:25074810, PMID:22292499, PMID:28298336). In cancer, GAPDH promotes NF-κB signaling via TRAF2 and drives a T-cell lymphoma phenotype when overexpressed (PMID:37450367, PMID:31447347).

Mechanistic history

Synthesis pass · year-by-year structured walk · 13 steps
  1. 1995 High

    Established that nitric oxide directly and reversibly inhibits GAPDH by modifying its active-site cysteine, defining the foundational redox-switch chemistry rather than ADP-ribosylation.

    Evidence In vitro assays with purified GAPDH and endothelial cells, stoichiometric nitrosylation quantification, substrate-protection and thiol-reversal controls

    PMID:7573405

    Open questions at the time
    • Did not connect cysteine nitrosylation to any downstream cellular function
    • No structural detail of the modified cysteine
  2. 2006 High

    Linked active-site S-nitrosylation to a cell-fate program by showing modified GAPDH binds Siah1 and translocates to the nucleus to trigger cytotoxicity, transforming GAPDH from a metabolic enzyme into an apoptotic signaling molecule.

    Evidence Co-IP, NOS inhibitor and binding-interference experiments, nuclear translocation assays, Cys150 mutagenesis

    PMID:16574384 PMID:16633896

    Open questions at the time
    • Nuclear effector targets of the GAPDH-Siah1 complex not yet defined
    • Physiological versus pathological triggers not separated
  3. 2008 High

    Defined the nuclear effector mechanism of translocated GAPDH (p300/CBP acetylation at Lys160 driving p53 apoptosis) and identified GAPDH as a redox-sensing component of a stress MAPK phosphorelay in yeast, generalizing the moonlighting paradigm.

    Evidence Co-IP, acetyltransferase activity assays, K160R dominant-negative mutagenesis; yeast genetic epistasis with Cys152 oxidation monitoring

    PMID:18406331 PMID:18552833

    Open questions at the time
    • Acetylation site mapping in vivo limited to single residue
    • Conservation of the yeast phosphorelay role in mammals not addressed
  4. 2009 High

    Identified GOSPEL as a cytosolic anchor competing with Siah1, establishing that nuclear translocation is actively regulated and providing a neuroprotective brake.

    Evidence Reciprocal Co-IP, competition binding, Cys47 mutagenesis, neuronal overexpression/knockdown, in vivo viral delivery

    PMID:19607794

    Open questions at the time
    • Structural basis of GOSPEL-GAPDH-Siah1 competition unresolved
    • GOSPEL's broader functions unknown
  5. 2012 Medium

    Extended GAPDH moonlighting to telomere biology and extracellular receptor functions, showing TERC binding inhibits telomerase and surface GAPDH acts as a plasminogen receptor for macrophage migration.

    Evidence Co-IP, TRAP telomerase assay, domain-mapping mutagenesis, senescence assays; surface binding assays and macrophage migration in vivo

    PMID:22847419 PMID:28298336

    Open questions at the time
    • Mechanism of GAPDH export to the cell surface unknown
    • TERC-binding interface mapped only to catalytic-domain lysine
  6. 2010 High

    Showed SNO-GAPDH transnitrosylates nuclear proteins (SIRT1, HDAC2, DNA-PK), providing a mechanism for delivering cytoplasmic NO signals to specific nuclear targets.

    Evidence In vitro transnitrosylation with purified proteins, cell-based Co-IP and translocation, Cys150 mutagenesis

    PMID:20972425

    Open questions at the time
    • Selectivity determinants for nuclear target choice unclear
    • Quantitative contribution versus direct NO nitrosylation not assessed
  7. 2014 Medium

    Resolved the apoptotic effector mechanism (GAPDH-p53 complex enhancing p53 expression/phosphorylation) and characterized GAPDH-AP-site Schiff-base adducts and iron-transport receptor activity, expanding the moonlighting catalog.

    Evidence Co-IP and interfering-peptide disruption with in vivo stroke model; NaBH4 crosslinking and MS; surface GAPDH iron-transport Co-IP and knockdown

    PMID:22292499 PMID:24670206 PMID:25074810 PMID:26203648

    Open questions at the time
    • GAPDH lacks AP lyase activity, so the functional role of AP-site adducts is undefined
    • Surface GAPDH iron-handling isoform differences not molecularly defined
  8. 2015 High

    Demonstrated that oxidative GAPDH inactivation is therapeutically exploitable, with high-dose vitamin C selectively killing glycolytic KRAS/BRAF-mutant colorectal cancer via ROS-mediated GAPDH inhibition.

    Evidence Cell viability and GAPDH activity assays, ROS/glutathione quantification, genetic mutant comparison, Apc/KrasG12D mouse model

    PMID:26541605

    Open questions at the time
    • Whether GAPDH is the sole oxidative-stress target driving death not fully isolated
    • Modification chemistry on the cysteine not site-mapped in this study
  9. 2018 High

    Established immunometabolic and oncogenic control of GAPDH activity through covalent succination (DMF) and arginine methylation (CARM1), and linked SNO-GAPDH/p300/SIRT1 signaling to tau acetylation in Alzheimer pathology.

    Evidence MS-confirmed Cys152 succination with flux analysis and in vivo immune readouts; in vitro methyltransferase and R234 mutant rescue with HCC samples; C150S mutant and CGP3466B rescue in Aβ mouse model and AD tissue

    PMID:29559585 PMID:29599194 PMID:30232003

    Open questions at the time
    • Interplay between competing modifications on the same enzyme not resolved
    • Relative contribution of glycolytic loss versus moonlighting effects in disease unclear
  10. 2019 High

    Provided the structural explanation for fumarate-derivative inhibition and added itaconate alkylation at Cys22 as an anti-inflammatory mechanism, while linking GAPDH overexpression to NF-κB-driven T-cell lymphoma.

    Evidence 2.29 Å crystal structure of MMF-Cys152 adduct with activity assay; 4-OI alkylation, U13C tracing and C22A rescue; T-cell GAPDH transgenic mouse with NIK inhibition

    PMID:31387164 PMID:31447347 PMID:31704924

    Open questions at the time
    • Cys22 is distinct from the catalytic Cys152, so distinct functional consequences need separation
    • Mechanism connecting GAPDH levels to non-canonical NF-κB not molecularly defined here
  11. 2020 High

    Defined GAPDH as a heme chaperone delivering heme to apo-sGCβ1 and as a nuclear partner of NAMPT for NAD+ salvage and DNA polymerase β for base excision repair, establishing chaperone and genome-maintenance moonlighting roles with defined regulatory inputs.

    Evidence In vitro heme transfer with purified proteins and live-cell reporter; SPR/SAXS/Co-IP for NAMPT complex; Src kinase assay and Tyr41 mutant with Pol β Co-IP and BER assay

    PMID:31988240 PMID:32358060 PMID:32539222

    Open questions at the time
    • Source and trafficking of GAPDH-bound heme within cells incompletely defined
    • How a single enzyme partitions among heme, NAD+ and BER roles is unknown
  12. 2021 Medium

    Quantified GAPDH's rate-controlling threshold in glycolysis and connected additional redox modifications (H2S sulfhydration, glutathionylation) and modification crosstalk to nuclear translocation, apoptosis, autophagy and aging phenotypes.

    Evidence siRNA/iodoacetate titration with metabolite and thermodynamic analysis; sulfhydration and glutathionylation Co-IP series with mutants and functional readouts; S-nitrosation proteomics in aged muscle

    PMID:33459133 PMID:33545174 PMID:34332079 PMID:34973445

    Open questions at the time
    • Whether distinct cysteine modifications produce identical or distinct nuclear outcomes is unresolved
    • Single-lab studies for several signaling axes lack reciprocal cross-validation
  13. 2023 Medium

    Established with a genetic knock-in that the GAPDH redox switch itself is required to route carbon to the oxidative PPP and support tumor growth, and identified serotonylation, acetate-driven acetylation, and VRK2/FBXW10/TRAF2 axes as activity-tuning inputs across immunity and cancer.

    Evidence Redox-switch knock-in mice/cells with flux analysis and tumor models; TGM2 serotonylation at Gln262 with CAR-T transfer; neutrophil NET assays; ubiquitination/phosphorylation biochemistry and NF-κB reporter with HCC mouse model

    PMID:37024754 PMID:37147288 PMID:37450367 PMID:38215751

    Open questions at the time
    • Integration of the many activity-tuning modifications into a unified regulatory logic is lacking
    • Acetylation site and acetyltransferase for the activating acetylation not identified [#31]

Open questions

Synthesis pass · forward-looking unresolved questions
  • How a single enzyme integrates competing cysteine and non-cysteine modifications to select among glycolytic, apoptotic, heme-chaperone, DNA-repair, and receptor functions, and what governs its localization to nucleus, cell surface, and extracellular space, remains unresolved.
  • No unified model of modification crosstalk and functional partitioning
  • Trafficking machinery for surface/extracellular GAPDH unknown
  • Structural basis for most moonlighting partner interactions undetermined

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0016491 oxidoreductase activity 4 GO:0140104 molecular carrier activity 3 GO:0140299 molecular sensor activity 3 GO:0001618 virus receptor activity 2 GO:0140096 catalytic activity, acting on a protein 2 GO:0003677 DNA binding 1 GO:0003723 RNA binding 1 GO:0140098 catalytic activity, acting on RNA 1
Localization
GO:0005634 nucleus 7 GO:0005829 cytosol 3 GO:0005886 plasma membrane 3 GO:0005576 extracellular region 1
Pathway
R-HSA-1430728 Metabolism 5 R-HSA-168256 Immune System 5 R-HSA-5357801 Programmed Cell Death 5 R-HSA-8953897 Cellular responses to stimuli 4 R-HSA-162582 Signal Transduction 3 R-HSA-73894 DNA Repair 2 R-HSA-8953854 Metabolism of RNA 1
Partners
EP300GOSPELNAMPTPOLBSIAH1SIRT1TERCTRAF2

Evidence

Reading pass · 37 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1995 Nitric oxide (via S-nitrosoglutathione) inhibits GAPDH by reversible S-nitrosylation of the active-site cysteine residue; inhibition required stoichiometric addition of ~1 mol NO per mol GAPDH monomer and was reversed by low-molecular-weight thiols, ruling out ADP-ribosylation as the primary mechanism. In vitro enzyme assay with purified GAPDH and endothelial cells; substrate-protection experiments; thiol-reversal assays; quantitative nitrosylation measurement The American Journal of Physiology High 7573405
2006 Diverse apoptotic stimuli activate iNOS/nNOS, leading to S-nitrosylation of GAPDH at Cys150 (rat), which abrogates catalytic activity and enables GAPDH to bind the E3-ubiquitin-ligase Siah1; the GAPDH–Siah1 complex translocates to the nucleus via Siah1's nuclear localization signal, where it stabilizes Siah1 and triggers cytotoxicity. Co-immunoprecipitation; NOS inhibitor and GAPDH-Siah binding interference experiments; nuclear translocation assays; apoptosis readouts Biochimica et Biophysica Acta / Cellular and Molecular Neurobiology (review of original work) High 16574384 16633896
2008 Nuclear GAPDH (translocated after S-nitrosylation) is acetylated at Lys160 by the acetyltransferase p300/CBP via direct protein interaction; this in turn stimulates p300/CBP catalytic activity, activating downstream targets including p53 to cause apoptosis. A K160R dominant-negative GAPDH mutant blocks p300/CBP activation and reduces cell death. Co-immunoprecipitation; acetyltransferase activity assays; dominant-negative GAPDH K160R mutagenesis; apoptosis gene expression and cell-death quantification Nature Cell Biology High 18552833
2008 In fission yeast, the GAPDH isoform Tdh1 physically associates with the Mcs4 response regulator and stress-responsive MAPKKKs; H2O2 stress transiently oxidizes Cys152 of Tdh1, enhancing its association with Mcs4 and enabling phosphorelay signaling to the MAP kinase cascade, placing GAPDH as an essential redox-sensing component of the peroxide-stress phosphorelay. Co-immunoprecipitation; genetic epistasis (Tdh1 deletion abolishes Mpr1-Mcs4 interaction and phosphorelay); Cys152 oxidation monitoring Molecular Cell High 18406331
2009 A cytosolic 52 kDa protein named GOSPEL physiologically binds GAPDH, competing with Siah1 for GAPDH binding and thereby retaining GAPDH in the cytosol and preventing nuclear translocation. S-nitrosylation of GOSPEL at Cys47 enhances GAPDH–GOSPEL binding and its neuroprotective effect; GOSPEL overexpression protects neurons from NMDA excitotoxicity while its depletion enhances death. Co-immunoprecipitation; competition binding assays (GOSPEL vs. Siah1 for GAPDH); GOSPEL overexpression and knockdown in primary neuron cultures; in vivo viral delivery in mice; S-nitrosylation site mutagenesis (C47) Neuron High 19607794
2010 S-nitrosylated GAPDH (SNO-GAPDH) translocates to the nucleus and transnitrosylates nuclear proteins including SIRT1, HDAC2, and DNA-PK, providing a mechanism for targeted nitrosylation of nuclear proteins by cytoplasmic NO sources. Co-immunoprecipitation; transnitrosylation assays with purified SNO-GAPDH and nuclear protein targets; cell-based nuclear translocation; site-directed mutagenesis at Cys150 Nature Cell Biology High 20972425
2015 High-dose vitamin C (as dehydroascorbate, DHA) is taken up by GLUT1, reduced intracellularly to vitamin C while depleting glutathione, causing ROS accumulation that inactivates GAPDH; this inhibition of GAPDH in highly glycolytic KRAS- or BRAF-mutant colorectal cancer cells causes an energetic crisis and selective cell death. Cellular viability assays; GAPDH activity measurement after DHA treatment; ROS and glutathione quantification; KRAS/BRAF mutant vs. wild-type comparison; in vivo Apc/KrasG12D mouse model Science High 26541605
2018 Dimethyl fumarate (DMF) succinates and covalently inactivates the catalytic cysteine (Cys152) of GAPDH both in vitro and in vivo in mice and humans; this inhibition down-regulates aerobic glycolysis in activated myeloid and lymphoid cells and mediates DMF's anti-inflammatory effects. In vitro GAPDH enzyme activity assay; mass spectrometry identification of succination at Cys152; cell metabolic flux analysis; in vivo mouse experiments; immune cell functional readouts Science High 29599194
2018 CARM1 (PRMT4) methylates GAPDH at Arg234, inhibiting its catalytic activity; glucose starvation upregulates CARM1, leading to hypermethylation and GAPDH inhibition. Re-expression of wild-type but not methylation-mimetic GAPDH restores glycolytic levels, and R234 hypomethylation is found in hepatocellular carcinoma relative to normal tissue. In vitro methyltransferase assay; GAPDH activity assay; R234 methylation-mimetic mutant rescue experiments; in vitro and in vivo tumor proliferation; clinical HCC sample analysis by MS Cell Reports High 30232003
2019 4-Octyl itaconate (4-OI) directly alkylates Cys22 of GAPDH, reducing its enzymatic activity; U13C glucose tracing confirmed blockade of glycolytic flux at GAPDH; anti-inflammatory effects of 4-OI in macrophages are abrogated by overexpression of C22A mutant GAPDH but rescued by wild-type GAPDH. In vitro GAPDH alkylation and activity assay; isotope-tracing metabolic flux analysis (U13C glucose); C22A GAPDH mutagenesis rescue; LPS-lethality in vivo model Nature Communications High 31704924
2012 GAPDH interacts with the telomerase RNA component (TERC) via its NAD+-binding Rossmann fold, inhibits telomerase activity, and induces telomere shortening and breast cancer cell senescence; a lysine residue in the catalytic domain is required for this inhibition; substrate G3P and the NO donor GSNO negatively regulate GAPDH inhibition of telomerase. Co-immunoprecipitation; telomerase activity assay (TRAP); domain-mapping mutagenesis; telomere length measurement; cellular senescence assay PNAS Medium 22847419
2014 Nuclear GAPDH (translocated via Siah1-dependent mechanism upon glutamate stimulation) forms a protein complex with p53, enhancing p53 expression and phosphorylation; disruption of GAPDH–p53 interaction by a peptide blocks glutamate-induced cell death and protects against ischemia-induced neuronal death in vivo in rats. Co-immunoprecipitation; interfering peptide disruption; p53 expression and phosphorylation assays; in vivo rat tMCAo stroke model Molecular Brain Medium 24670206
2020 GAPDH functions as a heme chaperone that delivers heme to apo-soluble guanylyl cyclase β1 (apo-sGCβ): apo-sGCβ associates with GAPDH in cells and dissociates upon heme binding; purified GAPDH-heme complex transfers heme to apo-sGCβ in vitro; heme delivery depends on cellular GAPDH expression levels and on GAPDH's ability to bind intracellular heme. Fluorescence quenching reporter (tetra-Cys sGCβ) in live cells; GAPDH knockdown/overexpression; GAPDH heme-binding mutant; Co-IP in cells; in vitro heme transfer with purified proteins The Journal of Biological Chemistry High 32358060
2020 GAPDH and NAMPT form a stable nuclear complex required for nuclear translocation of NAMPT; this translocation sustains the stress-induced NMN/NAD+ salvage pathway in the nucleus upon H2O2, GSNO, or DNA-damage stimuli. Immunoprecipitation; pulldown; surface plasmon resonance; immunofluorescence; SAXS; MS-based complex analysis; cellular NAD+ rescue experiments The Journal of Biological Chemistry High 31988240
2020 Src kinase phosphorylates GAPDH at Tyr41 under DNA damage stress; this phosphorylation is essential for GAPDH nuclear translocation and its DNA repair function. Nuclear GAPDH associates with DNA polymerase β (Pol β), promotes Pol β polymerase activity, and enhances base excision repair (BER) efficiency. Kinase assay; Tyr41 phosphorylation-deficient GAPDH mutant; nuclear fractionation; Co-IP with Pol β; BER activity assay; GAPDH knockdown with DNA damage sensitivity measurement in cells and xenografts FASEB Journal Medium 32539222
2021 Hydrogen sulfide (H2S) sulfhydrates the active-site cysteine of GAPDH, causing its nuclear redistribution; nuclear GAPDH interacts with CCAR2/DBC1, disrupting the inhibitory CCAR2–SIRT1 complex; activated SIRT1 then deacetylates LC3B to trigger autophagy flux and restrict intracellular Mycobacterium tuberculosis. Sulfhydration assay; nuclear fractionation; Co-IP (GAPDH–CCAR2, CCAR2–SIRT1); active-site cysteine GAPDH mutant; GAPDH ablation cells; LC3B deacetylation assay; intracellular mycobacterial growth assay Autophagy Medium 33459133
2021 H2O2-induced S-glutathionylation of GAPDH promotes its nuclear translocation; nuclear GAPDH forms a complex with SIRT1 and trans-glutathionylates SIRT1, inhibiting SIRT1 deacetylase activity; inactivated SIRT1 stably binds acetylated p53, initiating caspase-3 cleavage and apoptosis. Glutaredoxin-1 (Glrx) reverses GAPDH S-glutathionylation and prevents nuclear translocation. Co-IP; GAPDH redox-dead mutant (C150S/C152S); nuclear fractionation; SIRT1 deacetylase assay; caspase-3 cleavage assay; Glrx overexpression; human primary endothelial cells Free Radical Biology & Medicine Medium 34332079
2018 Amyloid-β1-42 exposure increases NO production leading to GAPDH S-nitrosylation (at Cys150); SNO-GAPDH activates p300 acetyltransferase, promotes SIRT1 nitrosylation/inactivation, and thereby increases tau acetylation at Lys280 and neurofibrillary tangle formation; GAPDH C150S mutant or the inhibitor CGP3466B abrogated Aβ-induced tau acetylation and cognitive impairment in mice. S-nitrosylation assays; p300 acetyltransferase assay; SIRT1 activity assay; tau acetylation MS; GAPDH C150S mutant; CGP3466B pharmacological inhibition; mouse behavioral tests; postmortem AD brain tissue Science Signaling High 29559585
2022 GAPDH (as heme chaperone) is involved in heme maturation of myoglobin and hemoglobins α, β, γ; GAPDH mutants H53A and K227A decrease heme content of these globins; GAPDH is found in complex with each globin and Hsp90 in cells; GAPDH knockdown in C2C12 myoblasts suppresses myoglobin heme maturation and in K562 erythroleukemia cells suppresses hemoglobin dimerization. GAPDH overexpression/knockdown; GAPDH mutant (H53A, K227A) rescue experiments; Co-IP (GAPDH-globin-Hsp90 complex); heme content measurement; differentiation models (C2C12, HiDEP-1, K562) FASEB Journal Medium 34972240
2016 δPKC phosphorylates GAPDH, decreasing GAPDH tetramerization and glycolytic activity; GAPDH oligomerization (tetramer formation) is required for its non-catalytic role in mitochondrial elimination under oxidative stress; a rationally designed pseudo-GAPDH (ψGAPDH) peptide inhibits δPKC-mediated GAPDH phosphorylation and GAPDH oligomerization, reducing mitochondrial elimination and increasing cardiac damage in a myocardial infarction model. δPKC kinase assay; GAPDH tetramerization assay; GAPDH glycolytic activity in vitro and ex vivo; mitochondrial elimination assay; cardiac ischemia-reperfusion injury animal model The Journal of Biological Chemistry Medium 27129213
2023 Cells expressing a GAPDH active-site cysteine redox-switch mutant (cannot be oxidized by H2O2) retain glycolytic activity but fail to stimulate the oxidative pentose phosphate pathway and enhance reductive capacity upon oxidative stress; this impairs anchorage-independent growth, spheroid formation, and in vivo tumor growth; fatty acid metabolism in kidney and heart is altered in mice lacking the GAPDH redox switch. GAPDH redox-switch knock-in mutant mice and cell lines; metabolic flux analysis; anchorage-independent growth assays; in vivo tumor models; chemo/radiotherapy combination experiments Nature Metabolism High 37024754
2021 Quantitative knockdown/inhibition showed that glycolytic flux in cancer cells is unaffected until GAPDH activity falls below ~19% of normal; below this threshold, glycolysis decreases proportionally because substrate G3P accumulation can no longer thermodynamically compensate; GAPDH + PGK1 together operate near equilibrium and define the rate-controlling step. siRNA GAPDH knockdown; iodoacetate-dependent enzymatic inhibition; direct glycolysis rate measurement; G3P concentration measurement; thermodynamic analysis The Journal of Biological Chemistry Medium 33545174
2023 Inhibition of GAPDH in neutrophils blocks glycolysis and promotes pentose phosphate pathway activity, blunts the respiratory burst, increases intracellular pH, and is sufficient to cause neutrophil extracellular trap (NET) formation in a neutrophil-elastase-dependent manner; blocking the pH increase prevented NET formation and cell death, identifying GAPDH as an intrinsic suppressor of NET formation. GAPDH inhibition (pharmacological); metabolic flux assays; neutrophil extracellular trap assays; intracellular pH measurement; elastase inhibition; patient neutrophil metabolomics (COVID-19) Nature Communications Medium 37147288
2024 Tissue transglutaminase 2 (TGM2) transfers serotonin (5-HT) to GAPDH at Gln262, catalyzing GAPDH serotonylation; this modification supports cytoplasmic localization of GAPDH, induces a glycolytic metabolic shift, and promotes antitumor activity of CD8+ T cells; monoamine oxidase A (MAOA) degrades 5-HT and acts as an intrinsic negative regulator; overexpression of TPH1 in CAR-T cells increased antitumor responses. TGM2-catalyzed serotonylation assay; Gln262 site identification by MS; GAPDH localization by fractionation; metabolic flux assays; CAR-T cell adoptive transfer in vivo Molecular Cell Medium 38215751
2019 Crystal structure of monomethyl fumarate (MMF)-bound human GAPDH at 2.29 Å resolution showed that MMF is covalently linked to the catalytic Cys152; the adduct blocks NAD+ co-substrate binding via steric hindrance of the nicotinamide moiety, explaining GAPDH inhibition by fumarate derivatives. X-ray crystallography at 2.29 Å; GAPDH enzyme activity assay; structural comparison with NAD+-bound GAPDH Molecules and Cells High 31387164
2014 Cell-surface GAPDH on macrophages functions as a moonlighting receptor: under iron depletion it binds holotransferrin (and lactoferrin) for iron import; under iron excess, a different surface GAPDH isoform recruits apotransferrin in association with ferroportin to facilitate iron efflux; GAPDH knockdown abolishes these iron transport activities. Co-immunoprecipitation; immunofluorescence; surface GAPDH expression correlation with iron transport; GAPDH knockdown; in vivo rodent iron-overload model Journal of Cell Science / Biochemistry and Cell Biology Medium 22292499 25074810
2012 Cell-surface GAPDH on macrophages functions as a plasminogen receptor; upon inflammation, macrophages recruit GAPDH to their surface to capture plasminogen, enabling ECM proteolysis and macrophage migration; GAPDH knockdown and in vivo approaches confirmed this role. Biochemical binding assay; Co-immunoprecipitation; GAPDH knockdown; in vivo macrophage migration assay FASEB Journal (2017 paper citing 2012 mechanism) Medium 28298336
2023 FBXW10 promotes GAPDH polyubiquitination and activation; VRK2-dependent phosphorylation of GAPDH at Ser151 is required for its ubiquitination and activation by FBXW10; activated GAPDH interacts with TRAF2, upregulating canonical and non-canonical NF-κB pathways and increasing PD-L1 and AR-VRK2 expression in hepatocellular carcinoma. Ubiquitination assay; phosphorylation mutagenesis (Ser151); Co-IP (GAPDH-TRAF2); NF-κB reporter assay; in vivo transgenic HCC mouse model; koningic acid (GAPDH inhibitor) treatment Cell Reports Medium 37450367
2021 Age-associated sarcopenia correlates with increased iNOS-dependent GAPDH S-nitrosylation at Cys150, Cys154, and Cys245; nuclear translocation of S-nitrosated GAPDH contributes to apoptosis in aged muscle; iNOS inhibition (1400W) or GAPDH S-nitrosation site mutation alleviated apoptosis of C2C12 cells. Quantitative S-nitrosation proteomics; iNOS inhibitor experiments; GAPDH S-nitrosation site mutagenesis; apoptosis assay; mouse aging model Nitric Oxide: Biology and Chemistry Medium 34973445
2009 Extracellular GAPDH binds to immunoglobulin-like domains I–VI and fibronectin type III repeats 4–5 of the cell adhesion molecule L1; GAPDH-dependent phosphorylation of L1 (using ATP) promotes L1-mediated neurite outgrowth and L1-Fc bead aggregation, which are blocked by alkaline phosphatase or kinase inhibitor. Protein-protein binding assay (domain mapping); surface biotinylation; neurite outgrowth assay; GAPDH antibody inhibition; exogenous GAPDH addition; alkaline phosphatase and kinase inhibitor controls Molecular and Cellular Neurosciences Medium 19285135
2012 CIB1 depletion in neuroblastoma and breast cancer cells promotes non-apoptotic cell death requiring nuclear GAPDH accumulation; CIB1 loss disrupts PI3K/AKT and Ras/MEK/ERK pathways; AKT inhibition alone maximally induces GAPDH nuclear accumulation, whereas concurrent ERK inhibition causes DNA damage response and cell death; MEK/ERK inhibition alone does not affect GAPDH localization. CIB1 siRNA knockdown; nuclear fractionation; pharmacological pathway inhibition (PI3K, AKT, MEK/ERK); cell death and DNA damage assays Oncogene Medium 22964641
2023 Acetate increases acetyl-CoA levels in CD4+ T cells, potentiating GAPDH acetylation, which in turn enhances GAPDH enzymatic activity, aerobic glycolysis, and Th1 cell polarization; reducing acetyl-CoA via fatty acid oxidation inhibition decreases acetyl-GAPDH levels. Transcriptome profiling; GAPDH acetylation measurement; GAPDH activity assay; glycolysis measurement; Th1 polarization assay; acetyl-CoA manipulation Molecular Biology of the Cell Low 37133968
2005 Amyloid-β peptide exposure promotes nuclear accumulation of a disulfide-linked form of GAPDH, which becomes detergent-insoluble; disulfide bonding reduces GAPDH enzymatic activity; increased GAPDH disulfide bonding was observed in detergent-insoluble extracts from Alzheimer's disease patient and transgenic mouse brain tissue. Cell fractionation; GAPDH activity assay; immunofluorescence; Western blot for disulfide-bonded GAPDH in AD brain tissue and transgenic mice FASEB Journal Medium 16186172
2019 GAPDH overexpression in T cells activates the non-canonical NF-κB pathway in transgenic mice, leading to development of a peripheral Tfh-like lymphoma recapitulating human angioimmunoblastic T cell lymphoma (AITL); NIK inhibition targeting NF-κB signaling prolonged survival of AITL-bearing mice. T cell-specific GAPDH transgenic mouse model; NF-κB pathway analysis; histology and immunophenotyping; NIK inhibitor treatment Cancer Cell Medium 31447347
2011 GAPDH depletion in human lung carcinoma cells causes accelerated cellular senescence (proliferation arrest, morphology change, SA-β-galactosidase staining, upregulation of DEC1 and GLB1) via sustained AMPK activation through phosphorylation of its α subunit at Thr172, in the absence of DNA damage; this is dependent on compromised glycolysis and energy crisis, and occurs independently of LKB1. GAPDH siRNA knockdown; SA-β-galactosidase staining; AMPK Thr172 phosphorylation measurement; metabolic and genetic rescue experiments Biochemical and Biophysical Research Communications Medium 21749859
2015 GAPDH interacts with apurinic/apyrimidinic (AP) sites in DNA by forming Schiff-base intermediates (borohydride-trappable); the interaction depends on GAPDH SH-groups (disulfide reduction abolishes adduct formation); GAPDH does not exhibit AP lyase activity despite forming adducts. NaBH4 crosslinking; mass spectrometry peptide mapping; AP site binding assay with purified GAPDH; thiol reduction experiments Mutation Research Medium 26203648
2022 GAPDH delivers heme to IDO1 and TDO via direct interaction; overexpression of GAPDH increased heme delivery to apo-IDO1 and apo-TDO in cells; Hsp90 interacted with apo-IDO1 but not apo-TDO and was required only for IDO1 heme insertion, not TDO. GAPDH overexpression/knockdown in cells; Co-IP (GAPDH-IDO1, GAPDH-TDO, Hsp90 interactions); IDO1/TDO heme content measurement; heme-deficient HEK293T cell system Free Radical Biology & Medicine Medium 35051612

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2015 Vitamin C selectively kills KRAS and BRAF mutant colorectal cancer cells by targeting GAPDH. Science (New York, N.Y.) 725 26541605
2005 GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiological genomics 598 15769908
2018 Dimethyl fumarate targets GAPDH and aerobic glycolysis to modulate immunity. Science (New York, N.Y.) 549 29599194
2010 The diverse functions of GAPDH: views from different subcellular compartments. Cellular signalling 497 20727968
2019 4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to exert anti-inflammatory effects. Nature communications 357 31704924
2008 Nitric oxide-induced nuclear GAPDH activates p300/CBP and mediates apoptosis. Nature cell biology 346 18552833
2010 GAPDH mediates nitrosylation of nuclear proteins. Nature cell biology 329 20972425
2012 GAPDH: a common enzyme with uncommon functions. Clinical and experimental pharmacology & physiology 229 21895736
2009 Novel roles for GAPDH in cell death and carcinogenesis. Cell death and differentiation 229 19779498
2010 Oxidatively modified glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Alzheimer's disease: many pathways to neurodegeneration. Journal of Alzheimer's disease : JAD 209 20164570
2023 4-Octyl itaconate inhibits aerobic glycolysis by targeting GAPDH to promote cuproptosis in colorectal cancer. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 170 36706634
2006 GAPDH as a sensor of NO stress. Biochimica et biophysica acta 168 16574384
2015 Critical protein GAPDH and its regulatory mechanisms in cancer cells. Cancer biology & medicine 160 25859407
2015 Cytosolic thiol switches regulating basic cellular functions: GAPDH as an information hub? Biological chemistry 153 25581756
2006 Nitric oxide-GAPDH-Siah: a novel cell death cascade. Cellular and molecular neurobiology 148 16633896
2012 Subcellular dynamics of multifunctional protein regulation: mechanisms of GAPDH intracellular translocation. Journal of cellular biochemistry 146 22388977
2013 Plant cytoplasmic GAPDH: redox post-translational modifications and moonlighting properties. Frontiers in plant science 142 24282406
2018 CARM1 Methylates GAPDH to Regulate Glucose Metabolism and Is Suppressed in Liver Cancer. Cell reports 127 30232003
2005 Amyloid-beta induces disulfide bonding and aggregation of GAPDH in Alzheimer's disease. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 127 16186172
1995 S-nitrosoglutathione reversibly inhibits GAPDH by S-nitrosylation. The American journal of physiology 120 7573405
2009 GOSPEL: a neuroprotective protein that binds to GAPDH upon S-nitrosylation. Neuron 117 19607794
2012 Novel insight into the role of GAPDH playing in tumor. Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico 109 22911551
2023 The GAPDH redox switch safeguards reductive capacity and enables survival of stressed tumour cells. Nature metabolism 90 37024754
2024 A GAPDH serotonylation system couples CD8+ T cell glycolytic metabolism to antitumor immunity. Molecular cell 81 38215751
2013 Basic biology of GAPDH. Advances in experimental medicine and biology 81 22851445
2012 Housekeeping gene selection advisory: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin are targets of miR-644a. PloS one 75 23091630
2017 Role of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) in DNA Repair. Biochemistry. Biokhimiia 71 28601074
2019 Regulation of Autophagy by Nuclear GAPDH and Its Aggregates in Cancer and Neurodegenerative Disorders. International journal of molecular sciences 69 31027346
2012 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induces cancer cell senescence by interacting with telomerase RNA component. Proceedings of the National Academy of Sciences of the United States of America 69 22847419
2018 Pleiotropic effects of moonlighting glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in cancer progression, invasiveness, and metastases. Cancer metastasis reviews 67 30209795
2002 Differential expression of GAPDH and beta3-actin in growing collateral arteries. Molecular and cellular biochemistry 67 12190113
2012 The multifunctional glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a novel macrophage lactoferrin receptor. Biochemistry and cell biology = Biochimie et biologie cellulaire 63 22292499
2008 Glycolytic enzyme GAPDH promotes peroxide stress signaling through multistep phosphorelay to a MAPK cascade. Molecular cell 63 18406331
2020 The Writers, Readers, and Erasers in Redox Regulation of GAPDH. Antioxidants (Basel, Switzerland) 60 33339386
2009 Expression of CPEB, GAPDH and U6snRNA in cervical and ovarian tissue during cancer development. APMIS : acta pathologica, microbiologica, et immunologica Scandinavica 60 19161537
2019 GAPDH Expression Predicts the Response to R-CHOP, the Tumor Metabolic Status, and the Response of DLBCL Patients to Metabolic Inhibitors. Cell metabolism 58 30827861
2023 GAPDH: A common housekeeping gene with an oncogenic role in pan-cancer. Computational and structural biotechnology journal 57 37664172
2020 GAPDH delivers heme to soluble guanylyl cyclase. The Journal of biological chemistry 54 32358060
2021 Hydrogen sulfide-induced GAPDH sulfhydration disrupts the CCAR2-SIRT1 interaction to initiate autophagy. Autophagy 53 33459133
2018 GAPDH as a model non-canonical AU-rich RNA binding protein. Seminars in cell & developmental biology 53 29574117
2014 Disruption of the nuclear p53-GAPDH complex protects against ischemia-induced neuronal damage. Molecular brain 52 24670206
2019 GAPDH Overexpression in the T Cell Lineage Promotes Angioimmunoblastic T Cell Lymphoma through an NF-κB-Dependent Mechanism. Cancer cell 49 31447347
2012 Human and pneumococcal cell surface glyceraldehyde-3-phosphate dehydrogenase (GAPDH) proteins are both ligands of human C1q protein. The Journal of biological chemistry 48 23086952
2014 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Alzheimer's disease. Pathologie-biologie 46 25246025
2007 GAPDH is not regulated in human glioblastoma under hypoxic conditions. BMC molecular biology 44 17597534
2014 Protein moonlighting in iron metabolism: glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Biochemical Society transactions 41 25399609
2011 Accelerated cellular senescence phenotype of GAPDH-depleted human lung carcinoma cells. Biochemical and biophysical research communications 41 21749859
2009 Extracellular GAPDH binds to L1 and enhances neurite outgrowth. Molecular and cellular neurosciences 41 19285135
2009 Lactobacillus plantarum 299v surface-bound GAPDH: a new insight into enzyme cell walls location. Journal of microbiology and biotechnology 41 20075631
2021 Oxidized GAPDH transfers S-glutathionylation to a nuclear protein Sirtuin-1 leading to apoptosis. Free radical biology & medicine 40 34332079
2013 GAPDH and intermediary metabolism. Advances in experimental medicine and biology 40 22851446
2009 The hepatitis delta virus RNA genome interacts with eEF1A1, p54(nrb), hnRNP-L, GAPDH and ASF/SF2. Virology 40 19464723
2022 GAPDH is involved in the heme-maturation of myoglobin and hemoglobin. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 39 34972240
1992 In situ localization of spermatogenic cell-specific glyceraldehyde 3-phosphate dehydrogenase (Gapd-s) messenger ribonucleic acid in mice. Biology of reproduction 39 1591341
2019 Dengue virus nonstructural 3 protein interacts directly with human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and reduces its glycolytic activity. Scientific reports 38 30804377
2018 Nitrosylation of GAPDH augments pathological tau acetylation upon exposure to amyloid-β. Science signaling 38 29559585
2014 Moonlighting cell-surface GAPDH recruits apotransferrin to effect iron egress from mammalian cells. Journal of cell science 38 25074810
2012 CIB1 prevents nuclear GAPDH accumulation and non-apoptotic tumor cell death via AKT and ERK signaling. Oncogene 38 22964641
2023 Neutrophil metabolomics in severe COVID-19 reveal GAPDH as a suppressor of neutrophil extracellular trap formation. Nature communications 37 37147288
2017 Moonlighting glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH): an evolutionarily conserved plasminogen receptor on mammalian cells. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 35 28298336
2015 Elevated GAPDH expression is associated with the proliferation and invasion of lung and esophageal squamous cell carcinomas. Proteomics 34 25944651
2020 A nicotinamide phosphoribosyltransferase-GAPDH interaction sustains the stress-induced NMN/NAD+ salvage pathway in the nucleus. The Journal of biological chemistry 30 31988240
2016 Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) Protein-Protein Interaction Inhibitor Reveals a Non-catalytic Role for GAPDH Oligomerization in Cell Death. The Journal of biological chemistry 30 27129213
2016 Cloning, expression and characterization of a mucin-binding GAPDH from Lactobacillus acidophilus. International journal of biological macromolecules 30 27180300
2000 Relative quantitative RT-PCR protocol for TrkB expression in neuroblastoma using GAPD as an internal control. BioTechniques 30 10769746
2021 Extracellular GAPDH Promotes Alzheimer Disease Progression by Enhancing Amyloid-β Aggregation and Cytotoxicity. Aging and disease 29 34341704
2020 Identification of novel biomarkers, MUC5AC, MUC1, KRT7, GAPDH, CD44 for gastric cancer. Medical oncology (Northwood, London, England) 29 32219571
2005 Cloning, characterization and DNA immunization of an Onchocerca volvulus glyceraldehyde-3-phosphate dehydrogenase (Ov-GAPDH). Biochimica et biophysica acta 29 15955451
1995 Genomic organization of a mouse glyceraldehyde 3-phosphate dehydrogenase gene (Gapd-s) expressed in post-meiotic spermatogenic cells. Developmental genetics 29 7736666
2003 RT-PCR for the pseudogene-free amplification of the glyceraldehyde-3-phosphate dehydrogenase gene (gapd). Molecular and cellular probes 28 14580401
2017 Physiology, phylogeny, early evolution, and GAPDH. Protoplasma 27 28265765
2022 Indoleamine dioxygenase and tryptophan dioxygenase activities are regulated through GAPDH- and Hsp90-dependent control of their heme levels. Free radical biology & medicine 26 35051612
2021 Determining the quantitative relationship between glycolysis and GAPDH in cancer cells exhibiting the Warburg effect. The Journal of biological chemistry 26 33545174
2015 Small molecules preventing GAPDH aggregation are therapeutically applicable in cell and rat models of oxidative stress. Free radical biology & medicine 26 26748070
2020 Src-mediated phosphorylation of GAPDH regulates its nuclear localization and cellular response to DNA damage. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 25 32539222
2008 Expression and compartmentalisation of the glycolytic enzymes GAPDH and pyruvate kinase in boar spermatogenesis. Reproduction, fertility, and development 24 18671919
2001 Diplonemid glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and prokaryote-to-eukaryote lateral gene transfer. Protist 24 11693658
2015 Cell specific differences in the protein abundances of GAPDH and Na(+),K(+)-ATPase in skeletal muscle from aged individuals. Experimental gerontology 23 26747222
2023 Acetate regulates GAPDH acetylation and T helper 1 cell differentiation. Molecular biology of the cell 22 37133968
2015 Streptococcus pneumoniae GAPDH Is Released by Cell Lysis and Interacts with Peptidoglycan. PloS one 22 25927608
2013 GAPDH, as a virulence factor. Advances in experimental medicine and biology 22 22851449
2013 GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents. Molecular biology international 22 23738070
2023 GAPDH Is a Novel Ferroptosis-Related Marker and Correlates with Immune Microenvironment in Lung Adenocarcinoma. Metabolites 21 36837761
2015 GAPDH-knockdown reduce rotenone-induced H9C2 cells death via autophagy and anti-oxidative stress pathway. Toxicology letters 21 25725130
2006 Nuclear translocation and overexpression of GAPDH by the hyper-pressure in retinal ganglion cell. Biochemical and biophysical research communications 21 16469296
2015 Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with apurinic/apyrimidinic sites in DNA. Mutation research 20 26203648
2013 Compartmentation of GAPDH. Advances in experimental medicine and biology 20 22851447
2023 Elevated FBXW10 drives hepatocellular carcinoma tumorigenesis via AR-VRK2 phosphorylation-dependent GAPDH ubiquitination in male transgenic mice. Cell reports 19 37450367
2022 GAPDH mediates plant reovirus-induced incomplete autophagy for persistent viral infection in leafhopper vector. Autophagy 19 36036160
2020 Covalent inhibitors of GAPDH: From unspecific warheads to selective compounds. European journal of medicinal chemistry 19 32898762
2019 Structural Study of Monomethyl Fumarate-Bound Human GAPDH. Molecules and cells 19 31387164
2015 Blocking GluR2-GAPDH ameliorates experimental autoimmune encephalomyelitis. Annals of clinical and translational neurology 19 25909084
2023 Electrochemical Genosensing of Overexpressed GAPDH Transcripts in Breast Cancer Exosomes. Analytical chemistry 18 36683335
2022 3-Bromo-Isoxazoline Derivatives Inhibit GAPDH Enzyme in PDAC Cells Triggering Autophagy and Apoptotic Cell Death. Cancers 18 35804925
2022 New roles for GAPDH, Hsp90, and NO in regulating heme allocation and hemeprotein function in mammals. Biological chemistry 18 36152339
2021 Disruption of the Complex between GAPDH and Hsp70 Sensitizes C6 Glioblastoma Cells to Hypoxic Stress. International journal of molecular sciences 18 33546324
2008 GAPD and tubulin are suitable internal controls for qPCR analysis of oral squamous cell carcinoma cell lines. Oral oncology 18 18621570
2022 GAPDH in neuroblastoma: Functions in metabolism and survival. Frontiers in oncology 17 36267982
2021 GAPDH S-nitrosation contributes to age-related sarcopenia through mediating apoptosis. Nitric oxide : biology and chemistry 16 34973445
2010 Association and heterogeneity at the GAPDH locus in Alzheimer's disease. Neurobiology of aging 15 20864222

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