Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli B834(DE3) cells | Bacillus anthracis |
expression in Escherichia coli, wild-type and mutant enzymes C42S, and Y367F, Y425F, and Y367/Y425F | Bacillus anthracis |
Crystallization (Comment) | Organism |
---|---|
crystal structure at 2.30 A resolution. The structures of the NADH and NADPH complexes at ca. 2.3 A resolution reveal that a loop consisting of residues Glu180-Thr187 becomes ordered and changes conformation on NAD(P)H binding | Bacillus anthracis |
sitting drop vapor diffusion method, using 16-26% 2-methyl-2,4-pentanediol, 0.2 M magnesium acetate, and 0.1 M sodium cacodylate, pH 6.5, at 15°C | Bacillus anthracis |
Protein Variants | Comment | Organism |
---|---|---|
Y367/Y425F | inactive mutant enzyme | Bacillus anthracis |
Y367F | kcat for NADH is 3.9fold lower than wild-type value, kcat for NADPH is 5.6fold lower than wild-type value | Bacillus anthracis |
Y367F | the mutant is 18% as active as wild type enzyme | Bacillus anthracis |
Y367F/Y425F | inactive | Bacillus anthracis |
Y425F | kcat for NADH is 30fold lower than wild-type value, kcat for NADPH is 93fold lower than wild-type value | Bacillus anthracis |
Y425F | the mutant is 1% as active as wild type enzyme | Bacillus anthracis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
NADH | 25°C, wild-type enzyme | Bacillus anthracis | |
0.002 | - |
NADH | wild type enzyme, at 25°C | Bacillus anthracis | |
0.002 | - |
CoA-disulfide | 25°C, wild-type enzyme. Cosubstrate: NADH | Bacillus anthracis | |
0.003 | - |
NADPH | 25°C, wild-type enzyme | Bacillus anthracis | |
0.006 | - |
NADPH | wild type enzyme, at 25°C | Bacillus anthracis | |
0.006 | - |
CoA-disulfide | 25°C, wild-type enzyme, cosubstrate: NADPH | Bacillus anthracis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus anthracis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Bacillus anthracis |
Q-Sepharose column chromatography and Ni-NTA column chromatography | Bacillus anthracis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
CoA disulfide + NADH | - |
Bacillus anthracis | CoA + NAD+ | - |
? | |
CoA disulfide + NADPH | - |
Bacillus anthracis | CoA + NADP+ | - |
? | |
CoA-disulfide + NADH + H+ | Bacillus anthracis CoADR can use either pyridine nucleotide equally well | Bacillus anthracis | CoA + NAD+ | - |
? | |
CoA-disulfide + NADPH + H+ | Bacillus anthracis CoADR can use either pyridine nucleotide equally well | Bacillus anthracis | CoA + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BACoADR | - |
Bacillus anthracis |
CoADR | - |
Bacillus anthracis |
coenzyme A-disulfide reductase | - |
Bacillus anthracis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
CoA-disulfide | 25°C, mutant enzyme Y25F. Cosubstrate: NADH | Bacillus anthracis | |
0.3 | - |
NADPH | mutant enzyme Y425F, at 25°C | Bacillus anthracis | |
0.9 | - |
NADH | 25°C, mutant enzyme Y25F | Bacillus anthracis | |
0.9 | - |
NADH | mutant enzyme Y425F, at 25°C | Bacillus anthracis | |
2 | 8 | NADPH | 25°C, wild-type enzyme | Bacillus anthracis | |
2 | 8 | NADPH | wild type enzyme, at 25°C | Bacillus anthracis | |
5 | - |
CoA-disulfide | 25°C, mutant enzyme Y367F. Cosubstrate: NADH | Bacillus anthracis | |
5 | - |
NADPH | mutant enzyme Y367F, at 25°C | Bacillus anthracis | |
7 | - |
NADH | 25°C, mutant enzyme Y367F | Bacillus anthracis | |
7 | - |
NADH | mutant enzyme Y367F, at 25°C | Bacillus anthracis | |
27 | - |
NADH | 25°C, wild-type enzyme | Bacillus anthracis | |
27 | - |
NADH | wild type enzyme, at 25°C | Bacillus anthracis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Bacillus anthracis | |
NADH | - |
Bacillus anthracis | |
NADH | CoADR exhibits dual specificity with respect to the NAD(P)H substrate | Bacillus anthracis | |
NADPH | - |
Bacillus anthracis | |
NADPH | CoADR exhibits dual specificity with respect to the NAD(P)H substrate | Bacillus anthracis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00027 | - |
NADH | wild type enzyme, at 25°C | Bacillus anthracis | |
0.0093 | - |
NADPH | wild type enzyme, at 25°C | Bacillus anthracis |