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Literature summary for 2.7.2.11 extracted from

  • Perez-Arellano, I.; Rubio, V.; Cervera, J.
    Mapping active site residues in glutamate-5-kinase. The substrate glutamate and the feed-back inhibitor proline bind at overlapping sites (2006), FEBS Lett., 580, 6247-6253.
    View publication on PubMed

Application

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(Commentary)

Cloned (Comment) Organism
proB cloning into pET-22b to yield pGKE and overexpression in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

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

Inhibitors Comment Organism Structure
L-proline
-
Escherichia coli

KM Value [mM]

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

Organism UniProt Comment Textmining
Escherichia coli P0A7B5
-
-

Purification (Commentary)

Purification (Comment) Organism
to homogeneity Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamate
-
Escherichia coli ADP + L-glutamate 5-phosphate
-
?

Synonyms

Synonyms Comment Organism
G5K
-
Escherichia coli
glutamate-5-kinase
-
Escherichia coli

IC50 Value

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