Cloned (Comment) | Organism |
---|---|
expresion of His-tagged wild-type subunits GCLM and GCLC and expression of His-tagged mutants in Escherichia coli BL21(DE3) | Drosophila melanogaster |
Protein Variants | Comment | Organism |
---|---|---|
C139S/C267S | sequential site-directed mutagenesis, exchange of cysteine residues in the modifier subunit, the cysteine residues seem not to be involved in intersubunit disulfide bonding | Drosophila melanogaster |
C213S/C214S/C267S | sequential site-directed mutagenesis, exchange of cysteine residues in the modifier subunit, which can still associate with the catalytic subunit, but no longer form intersubunit disulfides, the enzyme activity is reduced but still higher than the activity of the catalytic subunit alone, the mutant is more sensitive to inhibition by GSH, human modifier subunit can complement the defect, the mutant strain shows a reduced GSH level | Drosophila melanogaster |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
GSH | feedback inhibition, the elimination of disulfide bridges between the subunits renders the enzyme more sensitive to inhibition | Drosophila melanogaster |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.91 | - |
L-glutamate | 25°C, recombinant His-tagged wild-type enzyme | Drosophila melanogaster | |
0.92 | - |
L-glutamate | 25°C, recombinant His-tagged wild-type catalytic subunit | Drosophila melanogaster | |
0.97 | - |
L-glutamate | 25°C, recombinant His-tagged wild-type catalytic subunit with recombinant His-tagged mutant modifier subunit C213S/C214S/C267S | Drosophila melanogaster |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
1 * 80000, about, catalytic subunit, + 1 * 28000, about, modifier subunit, SDS-PAGE | Drosophila melanogaster |
35000 | - |
x * 35000, mutant enzyme modifier subunit, SDS-PAGE | Drosophila melanogaster |
80000 | - |
1 * 80000, about, catalytic subunit, + 1 * 28000, about, modifier subunit, SDS-PAGE | Drosophila melanogaster |
140000 | - |
holoenzyme, gel filtration | Drosophila melanogaster |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-Glu + L-Cys | Drosophila melanogaster | the enzyme has key influence on glutathione homeostasis | ADP + phosphate + gamma-L-Glu-L-Cys | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Drosophila melanogaster | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant HIs-tagged wild-type and mutant enzymes | Drosophila melanogaster |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Drosophila melanogaster |
8.39 | - |
Drosophila catalytic subunit as a hybrid with the wild-type human modifier subunit, substrate L-alpha-aminobutyrate | Drosophila melanogaster |
8.45 | - |
wild-type holoenzyme | Drosophila melanogaster |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-Glu + L-Cys | - |
Drosophila melanogaster | ADP + phosphate + gamma-L-Glu-L-Cys | - |
? | |
ATP + L-Glu + L-Cys | the enzyme has key influence on glutathione homeostasis | Drosophila melanogaster | ADP + phosphate + gamma-L-Glu-L-Cys | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 1 * 80000, about, catalytic subunit, + 1 * 28000, about, modifier subunit, SDS-PAGE | Drosophila melanogaster |
monomer | x * 35000, mutant enzyme modifier subunit, SDS-PAGE | Drosophila melanogaster |
More | enzyme is build of 2 subunits, a modifier and a catalytic subunit, the modifier subunit DmGCLM possesses cysteine residues, Cyys213, Cys214, and Cys267, which can form covalent interactions with the catalytic subunit DmGCLC and modify its activity, the activity of the holoenzyme is enhanced compared to the catalytic subunit alone | Drosophila melanogaster |
Synonyms | Comment | Organism |
---|---|---|
GCL | - |
Drosophila melanogaster |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Drosophila melanogaster |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Drosophila melanogaster |