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Literature summary for 6.3.2.2 extracted from

  • Kelly, B.S.; Antholine, W.E.; Griffith, O.W.
    Escherichia coli gamma-glutamylcysteine synthetase. Two active site metal ions affect substrate and inhibitor binding (2002), J. Biol. Chem., 277, 50-58.
    View publication on PubMed

Inhibitors

Inhibitors Comment Organism Structure
L-buthionine-S-sulfoximine mechanism-based inhibitor, in contrary to the mammalian enzyme form, the Escherichia coli enzyme is inhibited more weakly and slowly in presence of Mg2+, replacement of the metal by Mn2+ leads to increased binding affinity and inactivation rate Escherichia coli
additional information no inhibition by L-buthionine-R-sulfoximine Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Km values for diverse substrates in presence of Mg2+, or Mn2+, or both, kinetics Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ 2 divalent metal ions per enzyme molecule are bound, can be replaced by Mn2+ and Mg2+, binding mechanism and kinetics, overview Escherichia coli
Mg2+ 2 divalent metal ions per enzyme molecule are bound, Mg2+ sharpens the substrate specificity, increases the resistance to L-buthionine-S,R-sulfoximine, can be replaced by Mn2+ and Cu2+, binding mechanism and kinetics, overview Escherichia coli
Mn2+ 2 divalent metal ions per enzyme molecule are bound, Mg2+ broadens the substrate specificity, decreases the resistance to L-buthionine-S,R-sulfoximine, can be replaced by Mg2+ and Cu2+, binding mechanism and kinetics, overview Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-Glu + L-Cys Escherichia coli first and rate-limiting step in the GSH biosynthesis ADP + phosphate + gamma-L-Glu-L-Cys
-
ir
ATP + L-Glu + L-Cys Escherichia coli JM109 first and rate-limiting step in the GSH biosynthesis ADP + phosphate + gamma-L-Glu-L-Cys
-
ir

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A6W9
-
-
Escherichia coli JM109 P0A6W9
-
-

Purification (Commentary)

Purification (Comment) Organism
from strain JM109, to homogeneity Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine reaction mechanism Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
51
-
purified enzyme Escherichia coli

Storage Stability

Storage Stability Organism
-20°C, purified enzyme, 25% glycerol, stable for several months Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + alpha-ethyl-L-glutamate + L-alpha-aminobutyrate
-
Escherichia coli ADP + phosphate + alpha-ethyl-L-glutamyl-L-alpha-aminobutyrate
-
ir
ATP + alpha-methyl-L-glutamate + L-alpha-aminobutyrate
-
Escherichia coli ADP + phosphate + alpha-methyl-L-glutamyl-L-alpha-aminobutyrate
-
ir
ATP + D-Glu + L-alpha-aminobutyrate
-
Escherichia coli ADP + phosphate + gamma-D-Glu-L-alpha-aminobutyrate
-
ir
ATP + L-Glu + allo-L-threonine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-allo-L-threonine
-
ir
ATP + L-Glu + beta-amino-iso-butyrate
-
Escherichia coli ADP + phosphate + gamma-L-Glu-beta-amino-iso-butyrate
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ir
ATP + L-Glu + beta-chloro-L-alanine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-beta-chloro-L-alanine
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ir
ATP + L-Glu + beta-cyano-L-alanine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-beta-cyano-L-alanine
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ir
ATP + L-Glu + Gly
-
Escherichia coli ADP + phosphate + gamma-L-Glu-Gly
-
ir
ATP + L-Glu + Gly
-
Escherichia coli JM109 ADP + phosphate + gamma-L-Glu-Gly
-
ir
ATP + L-Glu + L-alanine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-alanine
-
ir
ATP + L-Glu + L-alpha-aminobutyrate
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-alpha-aminobutyrate
-
ir
ATP + L-Glu + L-C-allylglycine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-C-allylglycine
-
ir
ATP + L-Glu + L-Cys
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-Cys
-
ir
ATP + L-Glu + L-Cys first and rate-limiting step in the GSH biosynthesis Escherichia coli ADP + phosphate + gamma-L-Glu-L-Cys
-
ir
ATP + L-Glu + L-Cys
-
Escherichia coli JM109 ADP + phosphate + gamma-L-Glu-L-Cys
-
ir
ATP + L-Glu + L-Cys first and rate-limiting step in the GSH biosynthesis Escherichia coli JM109 ADP + phosphate + gamma-L-Glu-L-Cys
-
ir
ATP + L-Glu + L-isoleucine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-isoleucine
-
ir
ATP + L-Glu + L-leucine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-leucine
-
ir
ATP + L-Glu + L-norleucine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-norleucine
-
ir
ATP + L-Glu + L-norvaline
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-norvaline
-
ir
ATP + L-Glu + L-norvaline
-
Escherichia coli JM109 ADP + phosphate + gamma-L-Glu-L-norvaline
-
ir
ATP + L-Glu + L-serine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-serine
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ir
ATP + L-Glu + L-threonine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-threonine
-
ir
ATP + L-Glu + L-valine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-valine
-
ir
ATP + L-Glu + O-methyl-DL-serine
-
Escherichia coli ADP + phosphate + gamma-L-Glu-O-methyl-DL-serine
-
ir
ATP + L-Glu + S-methyl-L-Cys
-
Escherichia coli ADP + phosphate + gamma-L-Glu-L-S-methyl-Cys
-
ir
ATP + N-methyl-L-glutamate + L-alpha-aminobutyrate
-
Escherichia coli ADP + phosphate + N-methyl-L-glutamyl-L-alpha-aminobutyrate
-
ir
additional information substrate specificity, poor substrates are beta-glutamate, (R,S)-beta-methyl-DL-glutamate, (R,S)-gamma-methyl-glutamate, L-aspartate, and DL-alpha-aminoadipate Escherichia coli ?
-
?
additional information substrate specificity, poor substrates are beta-glutamate, (R,S)-beta-methyl-DL-glutamate, (R,S)-gamma-methyl-glutamate, L-aspartate, and DL-alpha-aminoadipate Escherichia coli JM109 ?
-
?

Synonyms

Synonyms Comment Organism
gamma-GCS
-
Escherichia coli
gamma-Glutamylcysteine synthetase
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
ATP
-
Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
additional information inhibition kinetics Escherichia coli