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Literature summary for 6.3.1.2 extracted from

  • Kim, J.N.; Cann, I.K.; Mackie, R.I.
    Purification, characterization, and expression of multiple glutamine synthetases from Prevotella ruminicola 23 (2012), J. Bacteriol., 194, 176-184.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
potassium arsenate activates the gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola

Cloned(Commentary)

Cloned (Comment) Organism
genes GSI, GSIII-1, and GSIII-2, sequence comparisons and phylogenetic analysis, recombinant expression of His-tagged enzymes in Escherichia coli strain BL21 CodonPlus (DE3) RIL Prevotella ruminicola

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
ATP isozyme GSIII-1, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
0.07
-
ATP isozyme GSIII-2, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
0.43
-
NH3 isozyme GSIII-2, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola
0.45
-
ATP isozyme GSI, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
0.48
-
NH3 isozyme GSIII-1, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola
0.62
-
L-glutamine isozyme GSIII-2, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
1.04
-
hydroxylamine isozyme GSIII-1, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
1.3
-
L-glutamine isozyme GSIII-1, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
1.7 2 L-glutamate isozyme GSIII-2, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola
1.91
-
ATP isozyme GSIII-1, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola
1.92
-
hydroxylamine isozyme GSIII-2, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
1.93
-
L-glutamine isozyme GSI, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
2.65
-
ATP isozyme GSIII-2, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola
2.7
-
hydroxylamine isozyme GSI, pH 6.0, 37°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
8.58
-
L-glutamate isozyme GSIII-1, pH 7.5, 37°C, glutamine synthetase activity Prevotella ruminicola

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
Prevotella ruminicola 5737
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ required, optimal concentrations of Mn2+ ions for GSI, GSIII-1, and GSIII-2 activities are 0.25, 0.5, and 1 mM, respectively Prevotella ruminicola
additional information Mg2+, Cu2+, Co2+, or Ca2+ cannot substitute for Mn2+ Prevotella ruminicola

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56000
-
x * 56000, recombinant isozyme GSI Prevotella ruminicola
83000
-
12 * 83000, recombinant isozyme GSIII-1, SDS-PAGE Prevotella ruminicola
83000
-
2 * 83000, recombinant isozyme GSIII-1, SDS-PAGE Prevotella ruminicola
83000
-
3 * 83000, recombinant isozyme GSIII-2, SDS-PAGE Prevotella ruminicola

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-glutamate + NH3 Prevotella ruminicola
-
ADP + phosphate + L-glutamine
-
?
ATP + L-glutamate + NH3 Prevotella ruminicola 23
-
ADP + phosphate + L-glutamine
-
?

Organism

Organism UniProt Comment Textmining
Prevotella ruminicola
-
genes GSI, GSIII-1, and GSIII-2
-
Prevotella ruminicola 23
-
genes GSI, GSIII-1, and GSIII-2
-

Purification (Commentary)

Purification (Comment) Organism
recombinant HIs-tagged isozymes GSI, GSIII-1, and GSIII-2 from Escherichia coli strain BL21 CodonPlus (DE3) RIL by nickel affinity chromatograaphy Prevotella ruminicola

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-glutamate + NH3
-
Prevotella ruminicola ADP + phosphate + L-glutamine
-
?
ATP + L-glutamate + NH3 GSIII-1, and GSIII-2, no activity of GSI Prevotella ruminicola ADP + phosphate + L-glutamine
-
?
ATP + L-glutamate + NH3
-
Prevotella ruminicola 23 ADP + phosphate + L-glutamine
-
?
ATP + L-glutamate + NH3 GSIII-1, and GSIII-2, no activity of GSI Prevotella ruminicola 23 ADP + phosphate + L-glutamine
-
?
hydroxylamine + L-glutamine + ATP gamma-glutamylhydroxamate synthetase activity, GSI, GSIII-1, and GSIII-2 Prevotella ruminicola L-gamma-glutamyl-hydroxamate + ammonia + ADP
-
r
hydroxylamine + L-glutamine + ATP gamma-glutamylhydroxamate synthetase activity, GSI, GSIII-1, and GSIII-2 Prevotella ruminicola 23 L-gamma-glutamyl-hydroxamate + ammonia + ADP
-
r
additional information the isozymes show glutamine gamma-transferase activity, overview. Activities of the isozymes catalyzing ATP hydrolysis, overview. GSI has an about 100fold lower ATPase activity compared to ATPase activities of GSIII-1 and -2 Prevotella ruminicola ?
-
?
additional information the isozymes show glutamine gamma-transferase activity, overview. Activities of the isozymes catalyzing ATP hydrolysis, overview. GSI has an about 100fold lower ATPase activity compared to ATPase activities of GSIII-1 and -2 Prevotella ruminicola 23 ?
-
?

Subunits

Subunits Comment Organism
? x * 56000, recombinant isozyme GSI Prevotella ruminicola
dimer 2 * 83000, recombinant isozyme GSIII-1, SDS-PAGE Prevotella ruminicola
dodecamer 12 * 83000, recombinant isozyme GSIII-1, SDS-PAGE Prevotella ruminicola
trimer 3 * 83000, recombinant isozyme GSIII-2, SDS-PAGE Prevotella ruminicola

Synonyms

Synonyms Comment Organism
gamma-glutamyl transferase
-
Prevotella ruminicola
gamma-glutamylhydroxamate synthetase
-
Prevotella ruminicola
Glutamine synthetase
-
Prevotella ruminicola

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
35
-
isozyme GSI, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
37
-
isozymes GSIII-1 and GSIII-2, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
50 60 GSIII-1 and GSIII-2 glutamine synthetase activity Prevotella ruminicola

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 50 the enzyme activity is drastically decreased above 50°C, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.6
-
isozyme GSI, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
6
-
isozymes GSIII-1 and GSIII-2, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
6.8 7 GSIII-1 and GSIII-2 glutamine synthetase activity Prevotella ruminicola

pH Range

pH Minimum pH Maximum Comment Organism
5.2 5.6 little activity of GSI below pH 5.2 and above pH 5.6, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola
5.6 7.2 activity range, GSIII-1 and GSIII-2, gamma-glutamylhydroxamate synthetase activity Prevotella ruminicola

Cofactor

Cofactor Comment Organism Structure
ADP
-
Prevotella ruminicola
ATP
-
Prevotella ruminicola

Expression

Organism Comment Expression
Prevotella ruminicola isozyme GSIII-2 is significantly induced by high concentrations of ammonia with increases in measured glutamine synthetase activity up

General Information

General Information Comment Organism
physiological function glutamine synthetase plays a particularly important role in nitrogen metabolism and is the principal source of N for protein and nucleic acid synthesis Prevotella ruminicola