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Literature summary 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)

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

Protein Variants

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

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.1.2 0.05
-
ATP D51A mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.05
-
NH4+ wild type enzyme, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.07
-
ATP D51S mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.12
-
ATP D51N mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.21
-
ATP wild type enzyme, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.32
-
L-glutamate D51A mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.37
-
L-glutamate D51S mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.55
-
NH4+ D51E mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.85
-
L-glutamate D51N mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.92
-
NH4+ D51N mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 1.5
-
L-glutamate wild type enzyme, pH 7.0, 30°C Trichormus azollae
6.3.1.2 5.95
-
NH4+ D51S mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 10.6
-
ATP D51E mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 12.45
-
NH4+ D51A mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 16
-
L-glutamate D51E mutant, pH 7.0, 30°C Trichormus azollae

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
6.3.1.2 Trichormus azollae Q9RHZ1 cyanobacterium
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
6.3.1.2 GlnA
-
Trichormus azollae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.1.2 0.33
-
L-glutamate D51E mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 2.6
-
L-glutamate D51A mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 13.1
-
L-glutamate D51S mutant, pH 7.0, 30°C Trichormus azollae
6.3.1.2 42
-
L-glutamate wild type enzyme, pH 7.0, 30°C Trichormus azollae

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.1.2 0.0002
-
L-methionine-DL-sulfoximine wild-type enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.0003
-
Phosphinothricin wild-type enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
6.3.1.2 0.1
-
Phosphinothricin D51E mutant enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae
6.3.1.2 2.94
-
L-methionine-DL-sulfoximine D51E mutant enzyme, biosynthetic reaction, pH 7.0, 30°C Trichormus azollae