Cloned (Comment) | Organism |
---|---|
expression of the His-tagged wild-type enzyme and mutants in Spodoptera frugiperda Sf9 cells via baculovirus infection | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
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 |
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 |
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 |
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 |
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 |
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 |
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 |
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 | Comment | Organism | Structure |
---|---|---|---|
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 | |
dithiothreitol | inhibition of wild-type holoenzyme and C553G mutant holoenzyme, the latter is more sensitive | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.5 | - |
L-cysteine | recombinant mutant C553G holoenzyme, pH 8.0, 37°C | Homo sapiens | |
0.7 | - |
L-Glu | recombinant wild-type holoenzyme, pH 8.0, 37°C | Homo sapiens | |
0.8 | - |
L-cysteine | recombinant wild-type holoenzyme, pH 8.0, 37°C | Homo sapiens | |
0.9 | - |
L-Glu | recombinant mutant C553G holoenzyme, pH 8.0, 37°C | Homo sapiens | |
3.6 | - |
L-2-aminobutyrate | recombinant wild-type holoenzyme, pH 8.0, 37°C | Homo sapiens | |
5 | - |
L-2-aminobutyrate | recombinant mutant C553G holoenzyme, pH 8.0, 37°C | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
75000 | - |
2 * 75000, recombinant wild-type and mutant enzymes, SDS-PAGE | Homo sapiens |
114000 | - |
recombinant wild-type and mutant holoenzymes, gel filtration | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-Glu + L-Cys | Homo sapiens | rate-limiting step in glutathione biosynthesis | ADP + phosphate + gamma-L-Glu-L-Cys | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Sf9 insect cells | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-glutamate + L-cysteine = ADP + phosphate + gamma-L-glutamyl-L-cysteine | Cys553 is not the active site residue | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.12 | - |
purified recombinant mutant C249G catalytic subunit and mutant C295G catalytic subunit | Homo sapiens |
0.17 | - |
purified recombinant mutant C52G catalytic subunit and mutant C248G catalytic subunit | Homo sapiens |
0.38 | - |
purified recombinant mutant C605G catalytic subunit | Homo sapiens |
0.48 | - |
purified recombinant mutant C501G catalytic subunit | Homo sapiens |
0.63 | - |
purified recombinant mutant C491G catalytic subunit | Homo sapiens |
0.92 | - |
purified recombinant mutant C553G catalytic subunit | Homo sapiens |
1.15 | - |
purified recombinant wild-type catalytic subunit | Homo sapiens |
1.83 | - |
purified recombinant mutant C553G holoenzyme | Homo sapiens |
4.67 | - |
purified recombinant mutant C249G holoenzyme | Homo sapiens |
5.17 | - |
purified recombinant mutant C605G holoenzyme | Homo sapiens |
5.33 | - |
purified recombinant mutant C295G holoenzyme | Homo sapiens |
5.67 | - |
purified recombinant mutant C248G holoenzyme | Homo sapiens |
6.17 | - |
purified recombinant wild-type holoenzyme | Homo sapiens |
6.67 | - |
purified recombinant mutant C52G holoenzyme | Homo sapiens |
7 | - |
purified recombinant mutants C491G holoenzyme and C501G holoenzyme | Homo sapiens |
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 | 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 | - |
? | |
ATP + L-Glu + L-Cys | rate-limiting step in glutathione biosynthesis | Homo sapiens | ADP + phosphate + gamma-L-Glu-L-Cys | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 75000, recombinant wild-type and mutant enzymes, SDS-PAGE | Homo sapiens |
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 | Comment | Organism |
---|---|---|
GLCL | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |