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Literature summary for 1.1.1.261 extracted from

  • Hayashi, J.; Yamamoto, K.; Yoneda, K.; Ohshima, T.; Sakuraba, H.
    Unique coenzyme binding mode of hyperthermophilic archaeal sn-glycerol-1-phosphate dehydrogenase from Pyrobaculum calidifontis (2016), Proteins, 84, 1786-1796.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Pyrobaculum calidifontis

Crystallization (Commentary)

Crystallization (Comment) Organism
to 2.45 A resolution, one homohexamer per asymmetric unit, with presence of the bound cofactor NADPH in subunits D, E, and F. The phosphate group at C2' of the adenine ribose of NADPH is tightly held through the five biased hydrogen bonds with Ser40 and Thr42. The NADPH molecule is pushed away from the normal coenzyme binding site Pyrobaculum calidifontis

Protein Variants

Protein Variants Comment Organism
S40A/T42A increase in activity with NADPH Pyrobaculum calidifontis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.006
-
NADPH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis
0.031
-
NADPH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis
0.077
-
NADH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis
0.1
-
NADH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis
0.31
-
dihydroxyacetone phosphate wild-type, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
0.72
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
0.75
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis
1.1
-
dihydroxyacetone phosphate wild-type, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis

Organism

Organism UniProt Comment Textmining
Pyrobaculum calidifontis A3MTM6
-
-
Pyrobaculum calidifontis VA1 A3MTM6
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dihydroxyacetone phosphate + NADH + H+
-
Pyrobaculum calidifontis sn-glycerol-1-phosphate + NAD+
-
?
dihydroxyacetone phosphate + NADH + H+
-
Pyrobaculum calidifontis VA1 sn-glycerol-1-phosphate + NAD+
-
?
dihydroxyacetone phosphate + NADPH + H+
-
Pyrobaculum calidifontis sn-glycerol-1-phosphate + NADP+
-
?
dihydroxyacetone phosphate + NADPH + H+
-
Pyrobaculum calidifontis VA1 sn-glycerol-1-phosphate + NADP+
-
?

Synonyms

Synonyms Comment Organism
EgsA
-
Pyrobaculum calidifontis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.26
-
NADPH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis
0.3
-
dihydroxyacetone phosphate wild-type, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
5.22
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
7
-
NADPH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis
10.4
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis
10.4
-
NADH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis
10.4
-
dihydroxyacetone phosphate wild-type, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis
11
-
NADH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis

Cofactor

Cofactor Comment Organism Structure
NADH strong preference for NADH Pyrobaculum calidifontis
NADPH 2.4% of the reaction rate with NADH Pyrobaculum calidifontis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.96
-
dihydroxyacetone phosphate wild-type, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
7.29
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADPH, pH 7.0, 50°C Pyrobaculum calidifontis
8.71
-
NADPH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis
9.39
-
dihydroxyacetone phosphate wild-type, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis
14
-
dihydroxyacetone phosphate mutant S40A/T42A, cosubstrate NADH, pH 7.0, 50°C Pyrobaculum calidifontis
101
-
NADH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis
142
-
NADH wild-type, pH 7.0, 50°C Pyrobaculum calidifontis
1155
-
NADPH mutant S40A/T42A, pH 7.0, 50°C Pyrobaculum calidifontis