Activating Compound | Comment | Organism | Structure |
---|---|---|---|
glutaredoxin 1 | reactivation of GSSG-treated ICL in the presence of glutaredoxin 1 within 20 min | Chlamydomonas reinhardtii | |
additional information | no reactivation of GSSG-treated ICL in the presence of thioredoxin 1 | Chlamydomonas reinhardtii |
Cloned (Comment) | Organism |
---|---|
wild-type and mutants cloned by the pET-3c-HIS/Escherichia coli BL21 expression system | Chlamydomonas reinhardtii |
Protein Variants | Comment | Organism |
---|---|---|
C165S | behaves as the wild-type | Chlamydomonas reinhardtii |
C165S/C247S | is inactivated by 5 mM GSSG and reactivated by treatment with either 20 mM reduced dithiothreitol or reduced glutaredoxin 1 as the wild-type protein. Complete shift in molecular mass corresponding to one glutathione adduct after 5 h of incubation, but prolonged incubation does not induce any additional glutathionylation | Chlamydomonas reinhardtii |
C165S/C247S/C301S | is inactivated by 5 mM GSSG and reactivated by treatment with either 20 mM reduced dithiothreitol or reduced glutaredoxin 1 as the wild-type protein. Complete shift in molecular mass corresponding to one glutathione adduct after 5 h of incubation, but prolonged incubation does not induce any additional glutathionylation | Chlamydomonas reinhardtii |
C178S | is totally inactive | Chlamydomonas reinhardtii |
C247S | behaves as the wild-type. Complete shift in molecular mass corresponding to one glutathione adduct after 5 h of incubation, but prolonged incubation does not induce any additional glutathionylation | Chlamydomonas reinhardtii |
C301S | behaves as the wild-type | Chlamydomonas reinhardtii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
GSSG | ICL can be inactivated by glutathionylation and reactivated by glutaredoxin after reduced dithiothreitol treatment, whereas thioredoxin does not appear to regulate ICL activity | Chlamydomonas reinhardtii | |
H2O2 | incubation with 0.1 mM H2O2 and 0.5mM GSH for 30 min results in a decrease of protein activity comparable with that obtained in the presence of GSSG. The addition of reduced dithiothreitol provides full reactivation of enzyme activity. Exposure to 1 mM H2O2 for 30 min results in an almost complete inactivation of the enzyme, whereas in the presence of a 10fold lower concentration of H2O2 (0.1 mM), ICL retains 20% of total activity after 30 min of incubation. GSH, reacting with sulfenic acid, can protect the protein from H2O2-mediated irreversible inactivation by glutathionylation | Chlamydomonas reinhardtii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46830 | - |
mutant C247S mass spectrometry | Chlamydomonas reinhardtii |
46860 | - |
mutant C178S mass spectrometry | Chlamydomonas reinhardtii |
46870 | - |
mutant C178S mass spectrometry, after 1 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
46870 | - |
wild-type, mass spectrometry | Chlamydomonas reinhardtii |
46890 | - |
mutant C178S mass spectrometry, after 24 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
46890 | - |
mutant C178S mass spectrometry, after 5 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47120 | - |
mutant C247S mass spectrometry, after 1 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47140 | - |
mutant C247S mass spectrometry, after 24 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47160 | - |
mutant C247S mass spectrometry, after 5 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47170 | - |
wild-type, mass spectrometry, after 1 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47190 | - |
wild-type, mass spectrometry, after 5 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
47510 | - |
wild-type, mass spectrometry, after 24 h of GSSG-treated ICL | Chlamydomonas reinhardtii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Chlamydomonas reinhardtii | - |
- |
- |
Purification (Comment) | Organism |
---|---|
to homogeneity, by Ni2+ affinity chromatography | Chlamydomonas reinhardtii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
9.5 | - |
purified ICL | Chlamydomonas reinhardtii |
Storage Stability | Organism |
---|---|
-20°C | Chlamydomonas reinhardtii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate | - |
Chlamydomonas reinhardtii | succinate + glyoxylate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ICL | - |
Chlamydomonas reinhardtii |
isocitrate lyase | - |
Chlamydomonas reinhardtii |