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Literature summary for 1.2.1.9 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
D-fructose 6-phosphate a slight increase in enzyme activity is observed in the presence of D-fructose 6-phosphate and coenzyme A (1.1fold) Sulfurisphaera tokodaii
D-glucose 1-phosphate activity increases by 1.4fold with increasing D-glucose 1-phosphate concentration from 0 to 0.1 mM. The enzyme shows K-type allosterism in which the positive cooperativity is abolished with concomitant activation by glucose 1-phosphate Sulfurisphaera tokodaii
D-glucose 1-phosphate D-glucose 1-phosphate does not influence the affinity for DL-glyceraldehyde-3-phosphate; but increases the turnover severalfold Sulfurisphaera tokodaii
additional information ATP, ADP, AMP, D-glucose 6-phosphate and galactose 1-phosphate exhibit no effect on enzyme activity Sulfurisphaera tokodaii

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
A198S/S199I the catalytic efficiency with NADP+ decreases while that with NAD+ increases by 2.5fold. Substitutions reduces the NADP/NAD preference ratio by more than 50% Sulfurisphaera tokodaii
A198S/S199I the mutant shows reduced catalytic efficiency with NADP+ and increased catalytic efficiency with NAD+ as compared to the wild type enzyme Sulfurisphaera tokodaii
S199I the catalytic efficiency (kcat/Km) with NADP+ decreases by 0.5fold while that with NAD+ remains unchanged. Substitution reduces the NADP/NAD preference ratio by more than 50% Sulfurisphaera tokodaii
S199I the mutant shows reduced catalytic efficiency with NADP+ and increased catalytic efficiency with NAD+ as compared to the wild type enzyme Sulfurisphaera tokodaii

Inhibitors

Inhibitors Comment Organism Structure
additional information no effect: ADP, AMP, D-glucose 6-phosphate and D-galactose 1-phosphate Sulfurisphaera tokodaii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.022
-
NADP+ pH 8.0, 60┬░C, mutant enzyme A198S/S199I Sulfurisphaera tokodaii
0.0222
-
NADP+ mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.023
-
NADP+ pH 8.0, 60┬░C, wild-type enzyme Sulfurisphaera tokodaii
0.0231
-
NADP+ wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.026
-
NADP+ pH 8.0, 60┬░C, mutant enzyme S199I Sulfurisphaera tokodaii
0.0263
-
NADP+ mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.0901
-
NADP+ wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.0979
-
NADP+ mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.187
-
NADP+ mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
17.2
-
NAD+ mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
17.2
-
NAD+ pH 8.0, 60┬░C, mutant enzyme A198S/S199I Sulfurisphaera tokodaii
18.9
-
NAD+ wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
19.4
-
NAD+ mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
25.8
-
NAD+ mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
25.8
-
NAD+ pH 8.0, 60┬░C, mutant enzyme S199I Sulfurisphaera tokodaii
26.4
-
NAD+ pH 8.0, 60┬░C, wild-type enzyme Sulfurisphaera tokodaii
26.4
-
NAD+ wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
29.3
-
NAD+ mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56000
-
4 * 56000, SDS-PAGE Sulfurisphaera tokodaii
56400
-
4 * 56400, calculated from amino acid sequence Sulfurisphaera tokodaii
56400
-
4 * 56400, calculated from sequence Sulfurisphaera tokodaii
227000
-
gel filtration Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-glyceraldehyde 3-phosphate + NAD+ + H2O Sulfurisphaera tokodaii NAD+ is a poor co-substrate 3-phospho-D-glycerate + NADH + H+
-
?
D-glyceraldehyde 3-phosphate + NADP+ + H2O Sulfurisphaera tokodaii
-
3-phospho-D-glycerate + NADPH + H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O Sulfurisphaera tokodaii
-
3-phospho-D-glycerate + NADPH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O Sulfurisphaera tokodaii 7
-
3-phospho-D-glycerate + NADPH + 2 H+
-
ir

Organism

Organism UniProt Comment Textmining
Sulfurisphaera tokodaii
-
-
-
Sulfurisphaera tokodaii Q96XP0
-
-
Sulfurisphaera tokodaii 7 Q96XP0
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-glyceraldehyde 3-phosphate + NAD+ + H2O NAD+ is a poor co-substrate Sulfurisphaera tokodaii 3-phospho-D-glycerate + NADH + H+
-
?
D-glyceraldehyde 3-phosphate + NAD+ + H2O the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM Sulfurisphaera tokodaii 3-phospho-D-glycerate + NADH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NAD+ + H2O the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM Sulfurisphaera tokodaii 7 3-phospho-D-glycerate + NADH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Sulfurisphaera tokodaii 3-phospho-D-glycerate + NADPH + H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Sulfurisphaera tokodaii 3-phospho-D-glycerate + NADPH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM. K-type allosterism in which the positive cooperativity is abolished with concomitant activation by D-glucose 1-phosphate Sulfurisphaera tokodaii 3-phospho-D-glycerate + NADPH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O
-
Sulfurisphaera tokodaii 7 3-phospho-D-glycerate + NADPH + 2 H+
-
ir
D-glyceraldehyde 3-phosphate + NADP+ + H2O the enzyme prefers NADP+ to NAD+. NAD+ is a poor cosubstrate. The NAD+-dependent reaction of St-GAPN shows no saturation up to 20 mM. K-type allosterism in which the positive cooperativity is abolished with concomitant activation by D-glucose 1-phosphate Sulfurisphaera tokodaii 7 3-phospho-D-glycerate + NADPH + 2 H+
-
ir

Subunits

Subunits Comment Organism
homotetramer 4 * 56000, SDS-PAGE Sulfurisphaera tokodaii
homotetramer 4 * 56400, calculated from amino acid sequence Sulfurisphaera tokodaii
tetramer 4 * 56000, SDS-PAGE Sulfurisphaera tokodaii
tetramer 4 * 56400, calculated from sequence Sulfurisphaera tokodaii

Synonyms

Synonyms Comment Organism
GAP dehydrogenase
-
Sulfurisphaera tokodaii
GAPN
-
Sulfurisphaera tokodaii
non-phosphorylating GAP dehydrogenase
-
Sulfurisphaera tokodaii
St-GAPN
-
Sulfurisphaera tokodaii
STK_24770 locus name Sulfurisphaera tokodaii

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
60
-
assay at Sulfurisphaera tokodaii
85
-
-
Sulfurisphaera tokodaii

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
at 60┬░C Sulfurisphaera tokodaii
8
-
assay at Sulfurisphaera tokodaii

Cofactor

Cofactor Comment Organism Structure
NAD+ poor cofactor Sulfurisphaera tokodaii
NADP+ preferred cofactor Sulfurisphaera tokodaii
NADP+ the enzyme prefers NADP+ to NAD+. NAD+ is a poor co-substrate 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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.378
-
NAD+ pH 8.0, 60┬░C, wild-type enzyme Sulfurisphaera tokodaii
0.378
-
NAD+ wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.434
-
NAD+ mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.434
-
NAD+ pH 8.0, 60┬░C, mutant enzyme S199I Sulfurisphaera tokodaii
0.637
-
NAD+ wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
0.639
-
NAD+ mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
1
-
NAD+ mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
1
-
NAD+ pH 8.0, 60┬░C, mutant enzyme A198S/S199I Sulfurisphaera tokodaii
1.63
-
NAD+ mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
48.1
-
NADP+ mutant enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
48.1
-
NADP+ pH 8.0, 60┬░C, mutant enzyme S199I Sulfurisphaera tokodaii
61.5
-
NADP+ mutant enzyme A198S/S199I, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
61.5
-
NADP+ pH 8.0, 60┬░C, mutant enzyme A198S/S199I Sulfurisphaera tokodaii
71.4
-
NADP+ mutant enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
96.6
-
NADP+ pH 8.0, 60┬░C, wild-type enzyme Sulfurisphaera tokodaii
96.6
-
NADP+ wild type enzyme, in the absence of D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
188
-
NADP+ mutant enzyme A198S/S199I, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii
203
-
NADP+ wild type enzyme, in the presence of 0.01 mM D-glucose 1-phosphate, in 50 mM EPPS/NaOH (pH 8.0), at 60┬░C Sulfurisphaera tokodaii