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Literature summary for 6.3.1.2 extracted from

  • Wray, L.V.; Fisher, S.H.
    Functional roles of the conserved Glu304 loop of Bacillus subtilis glutamine synthetase (2010), J. Bacteriol., 192, 5018-5025.
    View publication on PubMedView publication on EuropePMC

Protein Variants

Protein Variants Comment Organism
A305G mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 16% of wild-type activity with L-glutamate, 2% of wild-type activity with L-glutamine Bacillus subtilis
E304D mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 37% of wild-type activity with L-glutamate, 8% of wild-type activity with L-glutamine Bacillus subtilis
E304H mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 43% of wild-type activity with L-glutamate, 5% of wild-type activity with L-glutamine Bacillus subtilis
G302A mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 80% of wild-type activity with L-glutamate, 58% of wild-type activity with L-glutamine Bacillus subtilis
P306A mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 160% of wild-type activity with L-glutamate, 130% of wild-type activity with L-glutamine Bacillus subtilis
V300A mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 76% of wild-type activity with L-glutamate, 18% of wild-type activity with L-glutamine Bacillus subtilis
Y303A mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 36% of wild-type activity with L-glutamate, 8% of wild-type activity with L-glutamine Bacillus subtilis
Y303H mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 150% of wild-type activity with L-glutamate, 100% of wild-type activity with L-glutamine Bacillus subtilis
Y303L mutation in the E304 flap, a flexible 7-residue loop over the entrance to the active site. 87% of wild-type activity with L-glutamate, 46% of wild-type activity with L-glutamine Bacillus subtilis

Inhibitors

Inhibitors Comment Organism Structure
AMP
-
Bacillus subtilis
glycine
-
Bacillus subtilis
L-glutamine feedback inhibition Bacillus subtilis
L-methionine-S-sulfoximine
-
Bacillus subtilis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
NH3 wild-type, 25°C, pH not specified in the publication Bacillus subtilis
0.68
-
hydroxylamine mutant E304H, 25°C, pH not specified in the publication Bacillus subtilis
0.74
-
hydroxylamine mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis
0.83
-
hydroxylamine wild-type, 25°C, pH not specified in the publication Bacillus subtilis
1.2
-
ATP mutant E304H, 25°C, pH not specified in the publication Bacillus subtilis
2.4
-
ATP wild-type, 25°C, pH not specified in the publication Bacillus subtilis
3.4
-
L-glutamate mutant E304H, 25°C, pH not specified in the publication Bacillus subtilis
5.3
-
ATP mutant G302A, 25°C, pH not specified in the publication Bacillus subtilis
9.4
-
ATP mutant Y303H, 25°C, pH not specified in the publication Bacillus subtilis
10
-
L-glutamate mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis
11
-
ATP mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis
27
-
L-glutamate wild-type, 25°C, pH not specified in the publication Bacillus subtilis
32
-
NH3 mutant E304H, 25°C, pH not specified in the publication Bacillus subtilis
59
-
L-glutamate mutant Y303H, 25°C, pH not specified in the publication Bacillus subtilis
84
-
L-glutamate mutant G302A, 25°C, pH not specified in the publication Bacillus subtilis
120
-
NH3 mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamate + NH3
-
Bacillus subtilis ADP + phosphate + L-glutamine
-
?
L-glutamine + hydroxylamine + ADP partial reverse reaction Bacillus subtilis gamma-glutamylhydroxamate + NH3 + ?
-
?

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.1
-
mutant Y303H, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.1
-
mutant Y303L, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.12
-
mutant V300A, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.13
-
wild-type, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.2
-
mutant P306A, 25°C, pH not specified in the publication Bacillus subtilis AMP
0.25
-
mutant P306A, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.28
-
mutant G302A, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
0.5
-
wild-type, 25°C, pH not specified in the publication Bacillus subtilis AMP
0.61
-
mutant Y303A, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
1
-
mutant Y303H, 25°C, pH not specified in the publication Bacillus subtilis AMP
1.3
-
mutant Y303A, 25°C, pH not specified in the publication Bacillus subtilis AMP
1.4
-
mutant V300A, 25°C, pH not specified in the publication Bacillus subtilis AMP
1.5
-
mutant P306A, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
1.5
-
mutant Y303L, 25°C, pH not specified in the publication Bacillus subtilis AMP
1.7
-
mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis AMP
1.7
-
mutant G302A, 25°C, pH not specified in the publication Bacillus subtilis AMP
2.1
-
mutant A305G, 25°C, pH not specified in the publication Bacillus subtilis AMP
2.4
-
wild-type, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
2.7
-
mutant A305G, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
4.1
-
mutant Y303H, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
7
-
mutant V300A, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
11
-
mutant G302A, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
13
-
mutant Y303A, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
19
-
mutant Y303L, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
20
-
mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
26
-
mutant E304A, 25°C, pH not specified in the publication Bacillus subtilis L-methionine-S-sulfoximine
30
-
mutant E304A, 25°C, pH not specified in the publication Bacillus subtilis AMP
31
-
wild-type, 25°C, pH not specified in the publication Bacillus subtilis glycine
79
-
mutant E304D, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
140
-
mutant A305G, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
140
-
mutant E304A, 25°C, pH not specified in the publication Bacillus subtilis L-glutamine
350
-
mutant A305G, 25°C, pH not specified in the publication Bacillus subtilis glycine