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Literature summary for 1.2.1.13 extracted from

  • Ito, F.; Chishiki, H.; Fushinobu, S.; Wakagi, T.
    Comparative analysis of two glyceraldehyde-3-phosphate dehydrogenases from a thermoacidophilic archaeon, Sulfolobus tokodaii (2012), FEBS Lett., 586, 3097-3103.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information no influence on activity is observed on assaying of the enzyme with 5 mM substrate and 0.5 mM co-substrate with 1 mM ATP, ADP, AMP, D-glucose 6-phosphate, D-fructose 6-phosphate, galactose 1-phosphate, D-glucose 1-phosphate or coenzyme A in either the glycolytic or gluconeogenic reaction Sulfurisphaera tokodaii

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli C43 (DE3) cells Sulfurisphaera tokodaii

Inhibitors

Inhibitors Comment Organism Structure
3-phospho-D-glyceroyl phosphate
-
Sulfurisphaera tokodaii
DL-glyceraldehyde 3-phosphate
-
Sulfurisphaera tokodaii
NAD+
-
Sulfurisphaera tokodaii
NADH strong inhibition with excess co-substrate s observed and no activity is observed with over 2 mM NADH Sulfurisphaera tokodaii
NADP+
-
Sulfurisphaera tokodaii
NADPH strong inhibition with excess co-substrate s observed and no activity is observed with over 2 mM NADPH Sulfurisphaera tokodaii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
NADPH wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii
0.21
-
DL-glyceraldehyde 3-phosphate wild type enzyme, with NADP+ as cosubstrate, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
0.28
-
NADP+ wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
0.61
-
NADH wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii
0.77
-
DL-glyceraldehyde 3-phosphate wild type enzyme, with NAD+ as cosubstrate, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
5.8
-
3-phospho-D-glyceroyl phosphate wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii
10.7
-
phosphate wild type enzyme, with NADP+ as cosubstrate, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
16.8
-
NAD+ wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37500
-
4 * 37500, calculated from amino acid sequence Sulfurisphaera tokodaii
38000
-
4 * 38000, SDS-PAGE Sulfurisphaera tokodaii
148000
-
gel filtration Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-phospho-D-glyceroyl phosphate + NADPH + H+ Sulfurisphaera tokodaii
-
DL-glyceraldehyde 3-phosphate + phosphate + NADP+
-
r
DL-glyceraldehyde 3-phosphate + phosphate + NAD+ Sulfurisphaera tokodaii
-
3-phospho-D-glyceroyl phosphate + NADH + H+
-
r
DL-glyceraldehyde 3-phosphate + phosphate + NADP+ Sulfurisphaera tokodaii
-
3-phospho-D-glyceroyl phosphate + NADPH + H+
-
r

Organism

Organism UniProt Comment Textmining
Sulfurisphaera tokodaii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography and Superdex 200 gel filtration Sulfurisphaera tokodaii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-phospho-D-glyceroyl phosphate + NADPH + H+
-
Sulfurisphaera tokodaii DL-glyceraldehyde 3-phosphate + phosphate + NADP+
-
r
DL-glyceraldehyde 3-phosphate + phosphate + NAD+
-
Sulfurisphaera tokodaii 3-phospho-D-glyceroyl phosphate + NADH + H+
-
r
DL-glyceraldehyde 3-phosphate + phosphate + NADP+
-
Sulfurisphaera tokodaii 3-phospho-D-glyceroyl phosphate + NADPH + H+
-
r

Subunits

Subunits Comment Organism
homotetramer 4 * 38000, SDS-PAGE Sulfurisphaera tokodaii
homotetramer 4 * 37500, calculated from amino acid sequence Sulfurisphaera tokodaii

Synonyms

Synonyms Comment Organism
GAPDH
-
Sulfurisphaera tokodaii
phosphorylating GAP dehydrogenase
-
Sulfurisphaera tokodaii

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
95
-
-
Sulfurisphaera tokodaii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.5
-
at 60°C Sulfurisphaera tokodaii

Cofactor

Cofactor Comment Organism Structure
NAD+ the highest activity is almost equal for NADP+ and NAD+, whereas the affinity for NADP+ is higher than that for NAD+ Sulfurisphaera tokodaii
NADP+ the highest activity is almost equal for NADP+ and NAD+, whereas the affinity for NADP+ is higher than that for NAD+ Sulfurisphaera tokodaii
NADPH in the gluconeogenic direction, the enzyme shows a strong preference for NADPH, and only negligible activity is detected with NADH Sulfurisphaera tokodaii

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.035
-
NADPH wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii
0.081
-
NADH wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii
0.75
-
NADP+ wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
2.8
-
DL-glyceraldehyde 3-phosphate wild type enzyme, with NAD+ as cosubstrate, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
10.7
-
NAD+ wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
13.1
-
DL-glyceraldehyde 3-phosphate wild type enzyme, with NADP+ as cosubstrate, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 60°C Sulfurisphaera tokodaii
49.1
-
3-phospho-D-glyceroyl phosphate wild type enzyme, in 50 mM EPPS/NaOH (pH 8.0), 20 mM potassium phosphate, at 50°C Sulfurisphaera tokodaii

Expression

Organism Comment Expression
Sulfurisphaera tokodaii ATP, ADP, AMP, D-glucose 6-phosphate and galactose 1-phosphate exhibit no effect on enzyme activity additional information
Sulfurisphaera tokodaii a slight increase in enzyme activity is observed in the presence of D-fructose 6-phosphate and coenzyme A (1.1fold) up