Application | Comment | Organism |
---|---|---|
additional information | G5K and the homologous acetylglutamate kinase closely resemble each other concerning substrate binding and catalysis, but that they have different mechanisms of feed-back control, roles of K10, K217 and T169 in catalysis and ATP binding and of D150 in orienting the catalytic lysines, roles of D148 and D150 in glutamate binding and of D148 and N149 in proline binding | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
proB cloning into pET-22b to yield pGKE and overexpression in Escherichia coli BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D148A | is active, 150fold increased proline requirement | Escherichia coli |
D148N | is active, 40fold increased proline requirement | Escherichia coli |
D150A | is not active | Escherichia coli |
D150N | is active | Escherichia coli |
D170A | activity is less than 1% of that of wild-type G5K | Escherichia coli |
D170N | is active | Escherichia coli |
G51A | is active | Escherichia coli |
K10A | activity is less than 1% of that of wild-type G5K | Escherichia coli |
K217A | activity is less than 1% of that of wild-type G5K, decreased proline requirement | Escherichia coli |
K217R | is active, decreased proline requirement | Escherichia coli |
M214A | is active | Escherichia coli |
N149A | activity is less than 1% of that of wild-type G5K, 14fold increased proline requirement | Escherichia coli |
T169A | is active, decreased proline requirement | Escherichia coli |
T169S | is active, decreased proline requirement | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-proline | - |
Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.8 | - |
ATP | mutant D170N | Escherichia coli | |
2 | - |
ATP | wild-type | Escherichia coli | |
2.3 | - |
ATP | mutant D148N | Escherichia coli | |
3.4 | - |
ATP | mutant D170A | Escherichia coli | |
4.2 | - |
ATP | mutant D150N | Escherichia coli | |
4.3 | - |
ATP | mutant G51A | Escherichia coli | |
4.5 | - |
ATP | mutant N149A | Escherichia coli | |
6.1 | - |
ATP | mutant K10A | Escherichia coli | |
6.1 | - |
ATP | mutant T169S | Escherichia coli | |
6.4 | - |
ATP | mutant M214A | Escherichia coli | |
6.5 | - |
ATP | mutant D148A | Escherichia coli | |
17.6 | - |
ATP | mutant K217R | Escherichia coli | |
18.4 | - |
ATP | mutant T169A | Escherichia coli | |
24.6 | - |
ATP | mutant K217A | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P0A7B5 | - |
- |
Purification (Comment) | Organism |
---|---|
to homogeneity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-glutamate | - |
Escherichia coli | ADP + L-glutamate 5-phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
G5K | - |
Escherichia coli |
glutamate-5-kinase | - |
Escherichia coli |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.011 | - |
mutant T169A | Escherichia coli | L-proline | |
0.016 | - |
mutant K217A | Escherichia coli | L-proline | |
0.02 | - |
mutant K217R | Escherichia coli | L-proline | |
0.021 | - |
mutant T169S | Escherichia coli | L-proline | |
0.15 | - |
wild-type | Escherichia coli | L-proline | |
0.165 | - |
mutant M214A | Escherichia coli | L-proline | |
0.17 | - |
mutant D170A | Escherichia coli | L-proline | |
0.176 | - |
mutant D150N | Escherichia coli | L-proline | |
0.22 | - |
mutant D170N | Escherichia coli | L-proline | |
0.26 | - |
mutant K10A | Escherichia coli | L-proline | |
0.51 | - |
mutant G51A | Escherichia coli | L-proline | |
2.16 | - |
mutant N149A | Escherichia coli | L-proline | |
5.9 | - |
mutant D148N | Escherichia coli | L-proline | |
21 | - |
mutant D148A | Escherichia coli | L-proline |