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Literature summary for 6.3.1.2 extracted from

  • Clemente, M.T.; Marquez, A.J.
    Site-directed mutagenesis of Glu-297 from the alpha-polypeptide of Phaseolus vulgaris glutamine synthetase alters kinetic and structural properties and confers resistance to L-methionine sulfoximine (1999), Plant Mol. Biol., 40, 835-845.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ET8894 Phaseolus vulgaris

Protein Variants

Protein Variants Comment Organism
E297A Km for ammonium strongly increased, minor effects on activity in forward reaction, reverse reaction activity strongly decreased, strong effect on binding behavior to a 2',5'-ADP-sepharose Phaseolus vulgaris

Inhibitors

Inhibitors Comment Organism Structure
L-methionine sulfoximine forward reaction: complete inhibition of wild type enzyme, only slight inhibition of mutant enzyme, reverse reaction: slight inhibition of mutant and wild-type enzyme Phaseolus vulgaris

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
ATP wild-type and mutant enzyme, pH 7.5, 37°C Phaseolus vulgaris
2
-
L-glutamate mutant enzyme, pH 7.5, 37°C Phaseolus vulgaris
7
-
L-glutamate wild-type enzyme, pH 7.5, 37°C Phaseolus vulgaris

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
SDS-PAGE Phaseolus vulgaris
350000
-
gel filtration Phaseolus vulgaris

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Phaseolus vulgaris
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant mutant and wild-type enzyme Phaseolus vulgaris

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + glutamate + NH4+
-
Phaseolus vulgaris ADP + phosphate + L-glutamine
-
r
ATP + glutamate + NH4+ central enzyme of nitrogen metabolism, ammonia assimilation Phaseolus vulgaris ADP + phosphate + L-glutamine
-
r

Subunits

Subunits Comment Organism
octamer 8 * 43000, subunits by SDS-PAGE, native mass by gel filtration Phaseolus vulgaris

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
forward reaction of wild-type and mutant enzyme, reverse reaction of wild-type enzyme Phaseolus vulgaris
45
-
reverse reaction of mutant enzyme Phaseolus vulgaris

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
wild-type and mutant enzyme, forward and reverse reaction Phaseolus vulgaris