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Literature summary extracted from

  • Gabellieri, E.; Rahuel-Clermont, S.; Branlant, G.; Strambini, G.B.
    Effects of NAD+ binding on the luminescence of tryptophans 84 and 310 of glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus (1996), Biochemistry, 35, 12549-12559.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.12 expression of wild-type and mutant enzyme W84F in Escherichia coli Geobacillus stearothermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.12 W84F slightly lower Km-values for NAD+ and glyceraldehyde 3-phosphate, slightly higher Km-value for phosphate. The construction of the mutant permitts the identification of the individual fluorescence and phosphorescence characteristics of the two Trp residues W84 and W310 in the native enzyme Geobacillus stearothermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.12 0.11
-
 NAD+ mutant enzyme W84F Geobacillus stearothermophilus
1.2.1.12 0.15
-
 NAD+ wild-type enzyme Geobacillus stearothermophilus
1.2.1.12 0.31
-
 D-glyceraldehyde 3-phosphate mutant enzyme W48F Geobacillus stearothermophilus
1.2.1.12 0.8
-
 D-glyceraldehyde 3-phosphate wild-type enzyme Geobacillus stearothermophilus
1.2.1.12 8.3
-
 phosphate wild-type enzyme Geobacillus stearothermophilus
1.2.1.12 9.6
-
 phosphate mutant enzyme W84F Geobacillus stearothermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.12 Geobacillus stearothermophilus
-
 wild-type and mutant W84F
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.12
-
 Geobacillus stearothermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.12 D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
 Geobacillus stearothermophilus 3-phospho-D-glyceroyl phosphate + NADH
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.12 46
-
 D-glyceraldehyde 3-phosphate mutant enzyme W84F Geobacillus stearothermophilus
1.2.1.12 70
-
 D-glyceraldehyde 3-phosphate
-
 Geobacillus stearothermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.12 NAD+ effects of NAD+ binding on the luminescence of Trp84 and Trp310. NAD+-induced conformational change is sequential and subtle rearrangement in the structure of unligated subunits might be responsible for the negative cooperative behavior of NAD+ binding Geobacillus stearothermophilus