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Literature summary for 2.7.2.8 extracted from

  • Xu, M.; Rao, Z.; Dou, W.; Jin, J.; Xu, Z.
    Site-directed mutagenesis studies on the L-arginine-binding sites of feedback inhibition in N-acetyl-L-glutamate kinase (NAGK) from Corynebacterium glutamicum (2012), Curr. Microbiol., 64, 164-172.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Corynebacterium glutamicum

Protein Variants

Protein Variants Comment Organism
DELTA2-15 Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 71fold increased compared to wild-type Corynebacterium glutamicum
DELTA2-19 Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 148fold increased compared to wild-type Corynebacterium glutamicum
DELTA2-29 Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 163fold increased compared to wild-type Corynebacterium glutamicum
DELTA25 Km (N-acetyl-L-glutamate) highly increased compared to wild-type, kcat highly decreased compared to wild-type, IC50 (L-arginine) 17fold increased compared to wild-type Corynebacterium glutamicum
DELTA8 Km (N-acetyl-L-glutamate) increased compared to wild-type, kcat highly decreased compared to wild-type, IC50 (L-arginine) 16fold increased compared to wild-type Corynebacterium glutamicum
E19A Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 40fold increased compared to wild-type Corynebacterium glutamicum
E19R Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 61fold increased compared to wild-type Corynebacterium glutamicum
E281A Km (N-acetyl-L-glutamate) highly increased compared to wild-type, kcat similar to wild-type, IC50 (L-arginine) 21fold increased compared to wild-type Corynebacterium glutamicum
G287A Km (N-acetyl-L-glutamate) similar to wild-type, kcat decreased compared to wild-type, IC50 (L-arginine) 37fold increased compared to wild-type Corynebacterium glutamicum
G287D Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 40fold increased compared to wild-type Corynebacterium glutamicum
H268A Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 39fold increased compared to wild-type Corynebacterium glutamicum
H268N Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 58fold increased compared to wild-type Corynebacterium glutamicum
H26A Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 54fold increased compared to wild-type Corynebacterium glutamicum
R209A Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 46fold increased compared to wild-type Corynebacterium glutamicum
R209K Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 37fold increased compared to wild-type Corynebacterium glutamicum
W23A Km (N-acetyl-L-glutamate) similar to wild-type, kcat similar to wild-type, IC50 (L-arginine) 4.5fold increased compared to wild-type Corynebacterium glutamicum

Inhibitors

Inhibitors Comment Organism Structure
L-arginine
-
Corynebacterium glutamicum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.14
-
N-acetyl-L-glutamate mutant H26A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.25
-
N-acetyl-L-glutamate mutant H268N, pH and temperature not specified in the publication Corynebacterium glutamicum
3.32
-
N-acetyl-L-glutamate mutant G287D, pH and temperature not specified in the publication Corynebacterium glutamicum
3.43
-
N-acetyl-L-glutamate mutant E19R, pH and temperature not specified in the publication Corynebacterium glutamicum
3.52
-
N-acetyl-L-glutamate wild-type, pH and temperature not specified in the publication Corynebacterium glutamicum
3.61
-
N-acetyl-L-glutamate mutant E19A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.72
-
N-acetyl-L-glutamate mutant R209A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.76
-
N-acetyl-L-glutamate mutant H268A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.86
-
N-acetyl-L-glutamate mutant G287A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.89
-
N-acetyl-L-glutamate mutant W23A, pH and temperature not specified in the publication Corynebacterium glutamicum
3.96
-
N-acetyl-L-glutamate mutant R209K, pH and temperature not specified in the publication Corynebacterium glutamicum
6.98
-
N-acetyl-L-glutamate mutant DELTA8, pH and temperature not specified in the publication Corynebacterium glutamicum
7.84
-
N-acetyl-L-glutamate mutant DELTA2-15, pH and temperature not specified in the publication Corynebacterium glutamicum
8.23
-
N-acetyl-L-glutamate mutant DELTA2-19, pH and temperature not specified in the publication Corynebacterium glutamicum
8.45
-
N-acetyl-L-glutamate mutant DELTA2-29, pH and temperature not specified in the publication Corynebacterium glutamicum
13.68
-
N-acetyl-L-glutamate mutant E281A, pH and temperature not specified in the publication Corynebacterium glutamicum
25.74
-
N-acetyl-L-glutamate mutant DELTA25, pH and temperature not specified in the publication Corynebacterium glutamicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
SDS-PAGE Corynebacterium glutamicum

Organism

Organism UniProt Comment Textmining
Corynebacterium glutamicum
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Corynebacterium glutamicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + N-acetyl-L-glutamate
-
Corynebacterium glutamicum ADP + N-acetyl-L-glutamate 5-phosphate
-
?

Synonyms

Synonyms Comment Organism
argB
-
Corynebacterium glutamicum
NagK
-
Corynebacterium glutamicum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2
-
N-acetyl-L-glutamate mutant DELTA25, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
3.2
-
N-acetyl-L-glutamate mutant DELTA25, pH and temperature not specified in the publication Corynebacterium glutamicum
4.2
-
N-acetyl-L-glutamate mutant DELTA8, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
6.4
-
N-acetyl-L-glutamate mutant DELTA8, pH and temperature not specified in the publication Corynebacterium glutamicum
7.2
-
N-acetyl-L-glutamate mutant DELTA2-29, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
7.9
-
N-acetyl-L-glutamate mutant DELTA2-29, pH and temperature not specified in the publication Corynebacterium glutamicum
11.5
-
N-acetyl-L-glutamate mutant DELTA2-19, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
12.4
-
N-acetyl-L-glutamate mutant DELTA2-19, pH and temperature not specified in the publication Corynebacterium glutamicum
13.8
-
N-acetyl-L-glutamate mutant DELTA2-15, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
15.2
-
N-acetyl-L-glutamate mutant DELTA2-15, pH and temperature not specified in the publication Corynebacterium glutamicum
31.4
-
N-acetyl-L-glutamate wild-type, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
32.8
-
N-acetyl-L-glutamate mutant W23A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
34.9
-
N-acetyl-L-glutamate mutant E281A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
36.1
-
N-acetyl-L-glutamate mutant G287A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
36.7
-
N-acetyl-L-glutamate mutant G287A, pH and temperature not specified in the publication Corynebacterium glutamicum
37.4
-
N-acetyl-L-glutamate mutant R209K, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
38.3
-
N-acetyl-L-glutamate mutant G287D, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
38.5
-
N-acetyl-L-glutamate mutant E19A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
39.7
-
N-acetyl-L-glutamate mutant R209A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
39.7
-
N-acetyl-L-glutamate mutant W23A, pH and temperature not specified in the publication Corynebacterium glutamicum
41
-
N-acetyl-L-glutamate mutant E281A, pH and temperature not specified in the publication Corynebacterium glutamicum
41.1
-
N-acetyl-L-glutamate mutant H268A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
41.3
-
N-acetyl-L-glutamate mutant G287D, pH and temperature not specified in the publication Corynebacterium glutamicum
41.9
-
N-acetyl-L-glutamate mutant R209A, pH and temperature not specified in the publication Corynebacterium glutamicum
42.1
-
N-acetyl-L-glutamate mutant E19R, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
42.1
-
N-acetyl-L-glutamate mutant R209K, pH and temperature not specified in the publication Corynebacterium glutamicum
43
-
N-acetyl-L-glutamate mutant H26A, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
43.3
-
N-acetyl-L-glutamate mutant H268N, pH and temperature not specified in the publication Corynebacterium glutamicum
43.4
-
N-acetyl-L-glutamate mutant H26A, pH and temperature not specified in the publication Corynebacterium glutamicum
43.8
-
N-acetyl-L-glutamate mutant H268N, pH and temperature not specified in the publication, with 1 mM L-arginine Corynebacterium glutamicum
44.1
-
N-acetyl-L-glutamate mutant E19A, pH and temperature not specified in the publication Corynebacterium glutamicum
44.5
-
N-acetyl-L-glutamate mutant H268A, pH and temperature not specified in the publication Corynebacterium glutamicum
44.8
-
N-acetyl-L-glutamate mutant E19R, pH and temperature not specified in the publication Corynebacterium glutamicum
45.1
-
N-acetyl-L-glutamate wild-type, pH and temperature not specified in the publication Corynebacterium glutamicum

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.4
-
wild-type, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
1.8
-
mutant W23A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
6.4
-
mutant DELTA8, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
6.8
-
mutant DELTA25, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
8.6
-
mutant E281A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
14.8
-
mutant R209K, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
14.9
-
mutant G287A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
15.7
-
mutant H268A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
16.1
-
mutant E19A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
16.2
-
mutant G287D, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
18.5
-
mutant R209A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
21.7
-
mutant H26A, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
22.3
-
mutant H268N, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
24.4
-
mutant E19R, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
28.4
-
mutant DELTA2-15, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
59.2
-
mutant DELTA2-19, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine
65.2
-
mutant DELTA2-29, pH and temperature not specified in the publication Corynebacterium glutamicum L-arginine