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Literature summary for 1.2.1.12 extracted from

  • Bedhomme, M.; Adamo, M.; Marchand, C.H.; Couturier, J.; Rouhier, N.; Lemaire, S.D.; Zaffagnini, M.; Trost, P.
    Glutathionylation of cytosolic glyceraldehyde-3-phosphate dehydrogenase from the model plant Arabidopsis thaliana is reversed by both glutaredoxins and thioredoxins in vitro (2012), Biochem. J., 445, 337-347.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Arabidopsis thaliana

Protein Variants

Protein Variants Comment Organism
C149S the mutant shows 92% activity compared to the wild type enzyme Arabidopsis thaliana
C149S about 1% of wild-type activity, residue Cys149 is both essential for catalysis and the only accessible cysteine Arabidopsis thaliana
C153S the mutant shows 0.7% activity compared to the wild type enzyme Arabidopsis thaliana
C153S residue Cys153 has apparently no role in catalysis, in spite of the proximity in space with catalytic Cys149 Arabidopsis thaliana

Inhibitors

Inhibitors Comment Organism Structure
H2O2 inhibits enzyme activity by converting the thiolate of Cys149 into irreversibly oxidized forms, -SO2- and –SO3- via a labile sulfenate intermediate –SO?. Reduced glutathione prevents this irreversible process by reacting with Cys149 sulfenates to give rise to a mixed disulfide. Glutathionylated enzyme can be fully reactivated either by cytosolic glutaredoxin, via a glutathione-dependent monothiol mechanism, or, less efficiently, by cytosolic thioredoxins physiologically reduced by NADPH:thioredoxin reductase Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Arabidopsis thaliana 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
x * 40000, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+ Arabidopsis thaliana
-
3-phospho-D-glyceroyl phosphate + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
-
-
-
Arabidopsis thaliana P25858
-
-

Oxidation Stability

Oxidation Stability Organism
isoform GapC1 is sensitive to oxidation by 0.05 mM H2O2, which appears to inhibit enzyme activity by converting the thiolate of Cys149 (S) into irreversible oxidized forms (SO2 and SO3) via a labile sulfenate intermediate (SO) Arabidopsis thaliana

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.4
-
mutant enzyme C149S, 50 mM Tris/HCl (pH 7.5), at 25°C Arabidopsis thaliana
0.4
-
mutant C149S, pH 7.5, 25°C Arabidopsis thaliana
51.1
-
mutant enzyme C153S, 50 mM Tris/HCl (pH 7.5), at 25°C Arabidopsis thaliana
51.1
-
mutant C153S, pH 7.5, 25°C Arabidopsis thaliana
55.5
-
wild-type, pH 7.5, 25°C Arabidopsis thaliana
55.5
-
His-tagged wild type enzyme, 50 mM Tris/HCl (pH 7.5), at 25°C Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
Arabidopsis thaliana 3-phospho-D-glyceroyl phosphate + NADH
-
?
D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
Arabidopsis thaliana 3-phospho-D-glyceroyl phosphate + NADH + H+
-
?
additional information enzymatic activity strictly depends on residue Cys149. Catalytic Cys149 is the only solvent-exposed cysteine of the protein and its thiol is relatively acidic, pKa =5.7 Arabidopsis thaliana ?
-
?

Subunits

Subunits Comment Organism
? x * 40000, SDS-PAGE Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
At3g04120
-
Arabidopsis thaliana
GapC1
-
Arabidopsis thaliana
GapC1 isoform Arabidopsis thaliana
GAPDH
-
Arabidopsis thaliana

Cofactor

Cofactor Comment Organism Structure
NAD+ dependent on, the activity with NADPH is negligible (0.15% of the NADH-dependent specific activity) Arabidopsis thaliana