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Literature summary extracted from

  • Tu, Z.; Anders, M.W.
    Identification of an important cysteine residue in human glutamate-cysteine ligase catalytic subunit by site-directed mutagenesis (1998), Biochem. J., 336, 675-680.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.2 expression of the His-tagged wild-type enzyme and mutants in Spodoptera frugiperda Sf9 cells via baculovirus infection Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.2 C248G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens
6.3.2.2 C249G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, reduced activity of the holoenzyme compared to the wild-type enzyme Homo sapiens
6.3.2.2 C295G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens
6.3.2.2 C491G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens
6.3.2.2 C501G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens
6.3.2.2 C52G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens
6.3.2.2 C553G site-directed mutagenesis in the catalytic subunit, slightly reduced activity of the catalytic subunit, about 3.5fold reduced activity of the holoenzyme compared to the wild-type enzyme Homo sapiens
6.3.2.2 C605G site-directed mutagenesis in the catalytic subunit, reduced activity of the catalytic subunit, activity of the holoenzyme is similar to the wild-type enzyme Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.2.2 cysteamine inactivation of wild-type enzyme and mutant C553G after 90 min at 4°C, 0.2 mM cysteamine and 2 mM ATP Homo sapiens
6.3.2.2 dithiothreitol inhibition of wild-type holoenzyme and C553G mutant holoenzyme, the latter is more sensitive Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.2 0.5
-
L-cysteine recombinant mutant C553G holoenzyme, pH 8.0, 37°C Homo sapiens
6.3.2.2 0.7
-
L-Glu recombinant wild-type holoenzyme, pH 8.0, 37°C Homo sapiens
6.3.2.2 0.8
-
L-cysteine recombinant wild-type holoenzyme, pH 8.0, 37°C Homo sapiens
6.3.2.2 0.9
-
L-Glu recombinant mutant C553G holoenzyme, pH 8.0, 37°C Homo sapiens
6.3.2.2 3.6
-
L-2-aminobutyrate recombinant wild-type holoenzyme, pH 8.0, 37°C Homo sapiens
6.3.2.2 5
-
L-2-aminobutyrate recombinant mutant C553G holoenzyme, pH 8.0, 37°C Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.2.2 75000
-
2 * 75000, recombinant wild-type and mutant enzymes, SDS-PAGE Homo sapiens
6.3.2.2 114000
-
recombinant wild-type and mutant holoenzymes, gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.2 ATP + L-Glu + L-Cys Homo sapiens rate-limiting step in glutathione biosynthesis ADP + phosphate + gamma-L-Glu-L-Cys
-
?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.2 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.2 recombinant His-tagged wild-type and mutant enzymes from Sf9 insect cells Homo sapiens

Reaction

EC Number Reaction Comment Organism Reaction ID
6.3.2.2 ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine Cys553 is not the active site residue Homo sapiens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.2.2 0.12
-
purified recombinant mutant C249G catalytic subunit and mutant C295G catalytic subunit Homo sapiens
6.3.2.2 0.17
-
purified recombinant mutant C52G catalytic subunit and mutant C248G catalytic subunit Homo sapiens
6.3.2.2 0.38
-
purified recombinant mutant C605G catalytic subunit Homo sapiens
6.3.2.2 0.48
-
purified recombinant mutant C501G catalytic subunit Homo sapiens
6.3.2.2 0.63
-
purified recombinant mutant C491G catalytic subunit Homo sapiens
6.3.2.2 0.92
-
purified recombinant mutant C553G catalytic subunit Homo sapiens
6.3.2.2 1.15
-
purified recombinant wild-type catalytic subunit Homo sapiens
6.3.2.2 1.83
-
purified recombinant mutant C553G holoenzyme Homo sapiens
6.3.2.2 4.67
-
purified recombinant mutant C249G holoenzyme Homo sapiens
6.3.2.2 5.17
-
purified recombinant mutant C605G holoenzyme Homo sapiens
6.3.2.2 5.33
-
purified recombinant mutant C295G holoenzyme Homo sapiens
6.3.2.2 5.67
-
purified recombinant mutant C248G holoenzyme Homo sapiens
6.3.2.2 6.17
-
purified recombinant wild-type holoenzyme Homo sapiens
6.3.2.2 6.67
-
purified recombinant mutant C52G holoenzyme Homo sapiens
6.3.2.2 7
-
purified recombinant mutants C491G holoenzyme and C501G holoenzyme Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.2 ATP + L-Glu + L-2-aminobutyrate
-
Homo sapiens ADP + phosphate + L-Glu-2-aminobutyrate
-
?
6.3.2.2 ATP + L-Glu + L-Cys reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
?
6.3.2.2 ATP + L-Glu + L-Cys rate-limiting step in glutathione biosynthesis Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
?

Subunits

EC Number Subunits Comment Organism
6.3.2.2 dimer 2 * 75000, recombinant wild-type and mutant enzymes, SDS-PAGE Homo sapiens
6.3.2.2 More reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
6.3.2.2 GLCL
-
Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.2 37
-
assay at Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.2 8
-
assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.2 ATP
-
Homo sapiens