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Literature summary for 6.3.2.2 extracted from

  • Tu, Z.; Anders, M.W.
    Expression and characterization of human glutamate-cysteine ligase (1998), Arch. Biochem. Biophys., 354, 247-254.
    View publication on PubMed
Search for more literature for 6.3.2.2

Cloned(Commentary)

Cloned (Comment) Organism
coexpression of the His-tagged catalytic and regulatory subunits in Spodoptera frugiperda Sf9 cells via baculovirus infection, formation of the holoenzyme in the cells Homo sapiens

General Stability

General Stability Organism
activity of the holoenzyme and of the catalytic subunit is reduced by 20% and 10%, respectively, after 1 cycle of freezing and thawing Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
2-mercaptoethanol
-
 Homo sapiens
dithiothreitol
-
 Homo sapiens
GSH noncompetitive to L-glutamate, inhibition is not dependent on reduction of disulfide bonds between the 2 subunits in the holoenzyme Homo sapiens
L-buthionine sulfoximine 95% inhibition at 0.001 mM Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.5
-
 L-cysteine recombinant catalytic subunit, pH 8.0, 37°C Homo sapiens
0.7
-
 L-glutamate recombinant holoenzyme, pH 8.0, 37°C Homo sapiens
0.8
-
 L-cysteine recombinant holoenzyme, pH 8.0, 37°C Homo sapiens
1.7
-
 L-2-aminobutyrate recombinant catalytic subunit, pH 8.0, 37°C Homo sapiens
3.4
-
 L-2-aminobutyrate recombinant holoenzyme, pH 8.0, 37°C Homo sapiens
3.5
-
 L-glutamate recombinant catalytic subunit, pH 8.0, 37°C Homo sapiens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
114000
-
 recombinant holoenzyme, gel filtration Homo sapiens

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged holoenzyme and individual subunits from Sf9 insect cells Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 the activity of both the recombinant holoenzyme and catalytic subunit increased by 40% after removal of the His-tag Homo sapiens
1.15
-
 recombinant catalytic subunit Homo sapiens
6.17
-
 recombinant holoenzyme Homo sapiens

Storage Stability

Storage Stability Organism
4°C, purified holoenzyme, 10% loss of activity after 1 week Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-Glu + L-2-aminobutyrate
-
 Homo sapiens ADP + phosphate + L-Glu-2-aminobutyrate
-
 ?
ATP + L-Glu + L-Cys
-
 Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
 ?
ATP + L-Glu + L-Cys first and rate-limiting step in glutathione biosynthesis Homo sapiens ADP + phosphate + gamma-L-Glu-L-Cys
-
 ?

Subunits

Subunits Comment Organism
More reaction can be performed by the catalytic subunit alone, but presence of the regulatory subunit in the holoenzyme increases the activity and the specificity with L-2-aminobutyrate as substrate Homo sapiens

Synonyms

Synonyms Comment Organism
GLCL
-
 Homo sapiens

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.2
-
 GSH recombinant holoenzyme, pH 8.0, 37°C Homo sapiens
25.5
-
 GSH recombinant catalytic subunit, pH 8.0, 37°C Homo sapiens