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Literature summary for 6.3.1.2 extracted from

  • Crespo, J.L.; Guerrero, M.G.; Florencio, F.J.
    Mutational analysis of Asp51 of Anabaena azollae glutamine synthetase. D51E mutation confers resistance to the active site inhibitors L-methionine-DL-sulfoximine and phosphinothricin (1999), Eur. J. Biochem., 266, 1202-1209.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
mutant and native enzymes expressed in Escherichia coli ET6017 Trichormus azollae

Protein Variants

Protein Variants Comment Organism
D51A 7% of activity in forward reaction, 4% of activity in reverse reaction no effect on protein structure Trichormus azollae
D51E 5% of activity in forward reaction, 87% of activity in reverse reaction no effect on protein structure Trichormus azollae
D51N 1.7% of activity in forward reaction, 2.4% of activity in reverse reaction no effect on protein structure Trichormus azollae
D51R complete loss of activity, no effect on protein structure Trichormus azollae
D51S 24% of activity in forward reaction, 15% of activity in reverse reaction, no effect on protein structure Trichormus azollae

Inhibitors

Inhibitors Comment Organism Structure
L-methionine-DL-sulfoximine MSX, complete inactivation of wild-type and mutant enzymes except D51E mutant, which shows only 10% inhibition Trichormus azollae
Phosphinothricin PPT, complete inactivation of wild-type and mutant enzymes except D51E mutant, which shows 70% inhibition Trichormus azollae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05
-
ATP D51A mutant, pH 7.0, 30°C Trichormus azollae
0.05
-
NH4+ wild type enzyme, pH 7.0, 30°C Trichormus azollae
0.07
-
ATP D51S mutant, pH 7.0, 30°C Trichormus azollae
0.12
-
ATP D51N mutant, pH 7.0, 30°C Trichormus azollae
0.21
-
ATP wild type enzyme, pH 7.0, 30°C Trichormus azollae
0.32
-
L-glutamate D51A mutant, pH 7.0, 30°C Trichormus azollae
0.37
-
L-glutamate D51S mutant, pH 7.0, 30°C Trichormus azollae
0.55
-
NH4+ D51E mutant, pH 7.0, 30°C Trichormus azollae
0.85
-
L-glutamate D51N mutant, pH 7.0, 30°C Trichormus azollae
0.92
-
NH4+ D51N mutant, pH 7.0, 30°C Trichormus azollae
1.5
-
L-glutamate wild type enzyme, pH 7.0, 30°C Trichormus azollae
5.95
-
NH4+ D51S mutant, pH 7.0, 30°C Trichormus azollae
10.6
-
ATP D51E mutant, pH 7.0, 30°C Trichormus azollae
12.45
-
NH4+ D51A mutant, pH 7.0, 30°C Trichormus azollae
16
-
L-glutamate D51E mutant, pH 7.0, 30°C Trichormus azollae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + glutamate + NH4+ Trichormus azollae central enzyme of nitrogen metabolism ADP + phosphate + L-glutamine
-
r

Organism

Organism UniProt Comment Textmining
Trichormus azollae Q9RHZ1 cyanobacterium
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzymes and enzyme from host strain Trichormus azollae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + glutamate + NH4+
-
Trichormus azollae ADP + phosphate + L-glutamine
-
r
ATP + glutamate + NH4+ central enzyme of nitrogen metabolism Trichormus azollae ADP + phosphate + L-glutamine
-
r

Synonyms

Synonyms Comment Organism
GlnA
-
Trichormus azollae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.33
-
L-glutamate D51E mutant, pH 7.0, 30°C Trichormus azollae
2.6
-
L-glutamate D51A mutant, pH 7.0, 30°C Trichormus azollae
13.1
-
L-glutamate D51S mutant, pH 7.0, 30°C Trichormus azollae
42
-
L-glutamate wild type enzyme, pH 7.0, 30°C Trichormus azollae

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0002
-
L-methionine-DL-sulfoximine wild-type enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
0.0003
-
Phosphinothricin wild-type enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
0.1
-
Phosphinothricin D51E mutant enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
2.94
-
L-methionine-DL-sulfoximine D51E mutant enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae