Crystallization (Comment) | Organism |
---|---|
mutant D402P, to 2.6 A resolution, and comparison with wild-type. The C-terminal domain rotates by about 140 degrees between the two states that catalyze the adenylation and thioester-forming half-reactions. The domain rotation is accompanied by a change in the main chain torsional angles of residue D402 | Alcaligenes sp. AL3007 |
Protein Variants | Comment | Organism |
---|---|---|
D402A | 100fold decrease in ratio kcat/Km for CoA | Alcaligenes sp. AL3007 |
D402P | crystallization data. Mutant adopts the proposed adenylate-forming conformation with very little change to the overall structure.The ability of the mutant to catalyze the adenylate-forming half-reaction is reduced by about 3fold, catalysis of the second half-reaction is reduced by 4 orders of magnitude | Alcaligenes sp. AL3007 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0009 | - |
4-Chlorobenzoate | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.0012 | - |
4-Chlorobenzoate | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.0023 | - |
4-Chlorobenzoate | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.0131 | - |
ATP | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.0281 | - |
ATP | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.124 | - |
ATP | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.31 | - |
CoA | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
2 | - |
CoA | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
6 | - |
CoA | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alcaligenes sp. AL3007 | - |
- |
- |
Alcaligenes sp. AL3007 | Q8GN86 | mutant D402P | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-Chlorobenzoate + CoA + ATP | - |
Alcaligenes sp. AL3007 | 4-Chlorobenzoyl-CoA + AMP + diphosphate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.9 | - |
4-Chlorobenzoate | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
1 | - |
ATP | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
1.8 | - |
CoA | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
2.8 | - |
ATP | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
3.4 | - |
4-Chlorobenzoate | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
3.7 | - |
CoA | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
9.2 | - |
4-Chlorobenzoate | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
9.3 | - |
CoA | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
11.5 | - |
ATP | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
CoA | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
0.3 | - |
CoA | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
2 | - |
ATP | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
30 | - |
4-Chlorobenzoate | mutant D402P, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
30 | - |
CoA | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
40 | - |
ATP | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
90 | - |
ATP | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
400 | - |
4-Chlorobenzoate | mutant D402A, pH 7.5, 25°C | Alcaligenes sp. AL3007 | |
10000 | - |
4-Chlorobenzoate | wild-type, pH 7.5, 25°C | Alcaligenes sp. AL3007 |