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Literature summary for 1.2.1.12 extracted from

  • Roitel, O.; Ivinova, O.; Muronetz, V.; Nagradova, N.; Branlant, G.
    Thermal unfolding used as a probe to characterize the intra- and intersubunit stabilizing interactions in phosphorylating D-glyceraldehyde-3-phosphate dehydrogenase from Bacillus stearothermophilus (2002), Biochemistry, 41, 7556-7564.
    View publication on PubMed
Search for more literature for 1.2.1.12

Protein Variants

Protein Variants Comment Organism
D282G enzyme exists as dimer and tetramer, the tetramer is inactive, the dimer is slightly active, 650fold decrease in turnover number for NAD+, 5.8fold increase in Km-value for NAD+ compared to wild-type enzyme Geobacillus stearothermophilus
N313T mutant enzyme with a drastic decrease in thermostability, weakening of cooperative interactions between the catalytic and the cofactor domains and an inefficient binding of NAD+, mutant enzyme exists only as tetramer, 65fold decrease in turnover number for NAD+, 50fold increase in Km-value for NAD+ compared to wild-type enzyme Geobacillus stearothermophilus
T34Q/T39S/L43Q drastic decrease in thermostability, inefficient NAD+ binding, enzyme exists as dimer and tetramer, the tetramer is inactive, the dimer is slightly active, 650fold decrease in turnover number for NAD+, 4fold increase in Km-value for NAD+ compared to wild-type enzyme Geobacillus stearothermophilus
W310F mutant enzyme with a drastic decrease in thermostability, mutant enzyme exists only as tetramer, 2fold increase of Km-value for NAD+, 1.3fold decrease in turnover number compared to wild-type enzyme Geobacillus stearothermophilus
Y283V mutant enzyme with a drastic decrease in thermostability, dimeric form is inactive, KM-value and turnover-number of tetramer are nearly identical to that of the wild-type Geobacillus stearothermophilus
Y46G/R52G inactive mutant enzyme, only exists as dimer Geobacillus stearothermophilus
Y46G/S48G inactive mutant enzyme, only exists as dimer Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, wild-type Geobacillus stearothermophilus
0.06
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant Y283V Geobacillus stearothermophilus
0.1
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant W310F Geobacillus stearothermophilus
0.2
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant D282G Geobacillus stearothermophilus
0.29
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant T34Q/T39S/L43Q Geobacillus stearothermophilus
2.5
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant N313T Geobacillus stearothermophilus

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Substrates and Products (Substrate)

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
62
-
 maximum of thermal transition peak of tetrameric form of mutant apoenzyme N313T Geobacillus stearothermophilus
64.8
-
 maximum of thermal transition peak of dimeric mutant apoenzyme T34Q/T39S/L43Q Geobacillus stearothermophilus
64.8
-
 maximum of thermal transition peak of tetrameric mutant apoenzyme T34Q/T39S/L43Q Geobacillus stearothermophilus
65.4
-
 maximum of thermal transition peak of dimeric mutant apoenzyme Y46G/S48G Geobacillus stearothermophilus
65.5
-
 maximum of thermal transition peak of dimeric mutant apoenzyme Y46G/R52G Geobacillus stearothermophilus
68.4
-
 maximum of thermal transition peak of tetrameric mutant apoenzyme Y283V Geobacillus stearothermophilus
70.3
-
 maximum of thermal transition peak of tetrameric mutant apoenzyme D283G Geobacillus stearothermophilus
70.7
-
 maximum of thermal transition peak of tetrameric mutant apoenzyme W310F Geobacillus stearothermophilus
71
-
 maximum of thermal transition peak of dimeric mutant apoenzyme D282G Geobacillus stearothermophilus
73.5
-
 maximum of thermal transition peak of dimeric mutant apoenzyme Y283V Geobacillus stearothermophilus
78.5
-
 maximum of thermal transition peak of tetrameric form of wild-type apoenzyme Geobacillus stearothermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.1
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant D282G Geobacillus stearothermophilus
0.1
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant T34Q/T39S/L43Q Geobacillus stearothermophilus
1
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant N313T Geobacillus stearothermophilus
51
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant W310F Geobacillus stearothermophilus
65
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, wild-type Geobacillus stearothermophilus
73
-
 NAD+ pH 8.0, 25°C, tetrameric enzyme form, mutant Y283V Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Geobacillus stearothermophilus