Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Escherichia coli |
Crystallization (Comment) | Organism |
---|---|
molecular replacement with the apo-AlaRS structure from Aquifex aeolicus,Protein Data Bank accession code 1RIQ, as the initial model | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D235A | no improvement in the discrimination between alanine and serine | Escherichia coli |
D235E | no improvement in the discrimination between alanine and serine | Escherichia coli |
D235N | no improvement in the discrimination between alanine and serine | Escherichia coli |
D235Q | no improvement in the discrimination between alanine and serine | Escherichia coli |
G237A | mutation G237A introduces bulk into the alanine-binding pocket, with little other change in the pocket or the surrounding atoms. Shrinking of the alanine-binding pocket sharply raises the Km for alanine but does not greatly perturb the Km for serine | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.05 | - |
L-alanine | wild-type, 22°C, pH 8.0 | Escherichia coli | |
26 | - |
glycine | wild-type, 22°C, pH 8.0 | Escherichia coli | |
31 | - |
L-serine | wild-type, 22°C, pH 8.0 | Escherichia coli | |
42 | - |
glycine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
58 | - |
L-serine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
70 | - |
L-serine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
73 | - |
L-alanine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
88 | - |
glycine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
178 | - |
L-alanine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
493 | - |
glycine | mutant D235A, 22°C, pH 8.0 | Escherichia coli | |
502 | - |
L-alanine | mutant D235A, 22°C, pH 8.0 | Escherichia coli | |
600 | - |
L-serine | above 600, mutant D235A, 22°C, pH 8.0 | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00957 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + glycine + tRNAAla | - |
Escherichia coli | AMP + diphosphate + glycyl-tRNAAla | - |
? | |
ATP + L-alanine + tRNAAla | - |
Escherichia coli | AMP + diphosphate + L-alanyl-tRNAAla | - |
? | |
ATP + L-serine + tRNAAla | - |
Escherichia coli | AMP + diphosphate + L-seryl-tRNAAla | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0028 | - |
L-serine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
0.0035 | - |
glycine | mutant D235A, 22°C, pH 8.0 | Escherichia coli | |
0.0046 | - |
L-serine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
0.0058 | - |
glycine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
0.0074 | - |
L-alanine | mutant D235N, 22°C, pH 8.0 | Escherichia coli | |
0.069 | - |
L-alanine | mutant D235A, 22°C, pH 8.0 | Escherichia coli | |
1.2 | - |
L-alanine | wild-type, 22°C, pH 8.0 | Escherichia coli | |
1.4 | - |
L-alanine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
1.6 | - |
glycine | mutant G237A, 22°C, pH 8.0 | Escherichia coli | |
2.4 | - |
L-serine | wild-type, 22°C, pH 8.0 | Escherichia coli | |
3.7 | - |
glycine | wild-type, 22°C, pH 8.0 | Escherichia coli |