BRENDA - Enzyme Database

Regulation by glutathionylation of isocitrate lyase from Chlamydomonas reinhardtii

Bedhomme, M.; Zaffagnini, M.; Marchand, C.H.; Gao, X.H.; Moslonka-Lefebvre, M.; Michelet, L.; Decottignies, P.; Lemaire, S.D.; J. Biol. Chem. 284, 36282-36291 (2009)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
4.1.3.1
glutaredoxin 1
reactivation of GSSG-treated ICL in the presence of glutaredoxin 1 within 20 min
Chlamydomonas reinhardtii
4.1.3.1
additional information
no reactivation of GSSG-treated ICL in the presence of thioredoxin 1
Chlamydomonas reinhardtii
Cloned(Commentary)
EC Number
Commentary
Organism
4.1.3.1
wild-type and mutants cloned by the pET-3c-HIS/Escherichia coli BL21 expression system
Chlamydomonas reinhardtii
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.1.3.1
C165S
behaves as the wild-type
Chlamydomonas reinhardtii
4.1.3.1
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
4.1.3.1
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
4.1.3.1
C178S
is totally inactive
Chlamydomonas reinhardtii
4.1.3.1
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
4.1.3.1
C301S
behaves as the wild-type
Chlamydomonas reinhardtii
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.1.3.1
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
4.1.3.1
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]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.1.3.1
46830
-
mutant C247S mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46860
-
mutant C178S mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46870
-
mutant C178S mass spectrometry, after 1 h of GSSG-treated ICL; wild-type, mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46890
-
mutant C178S mass spectrometry, after 24 h of GSSG-treated ICL; mutant C178S mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47120
-
mutant C247S mass spectrometry, after 1 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47140
-
mutant C247S mass spectrometry, after 24 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47160
-
mutant C247S mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47170
-
wild-type, mass spectrometry, after 1 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47190
-
wild-type, mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47510
-
wild-type, mass spectrometry, after 24 h of GSSG-treated ICL
Chlamydomonas reinhardtii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.1.3.1
Chlamydomonas reinhardtii
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.1.3.1
to homogeneity, by Ni2+ affinity chromatography
Chlamydomonas reinhardtii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.1.3.1
9.5
-
purified ICL
Chlamydomonas reinhardtii
Storage Stability
EC Number
Storage Stability
Organism
4.1.3.1
-20°C
Chlamydomonas reinhardtii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.1.3.1
isocitrate
-
704605
Chlamydomonas reinhardtii
succinate + glyoxylate
-
-
-
?
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
4.1.3.1
glutaredoxin 1
reactivation of GSSG-treated ICL in the presence of glutaredoxin 1 within 20 min
Chlamydomonas reinhardtii
4.1.3.1
additional information
no reactivation of GSSG-treated ICL in the presence of thioredoxin 1
Chlamydomonas reinhardtii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.1.3.1
wild-type and mutants cloned by the pET-3c-HIS/Escherichia coli BL21 expression system
Chlamydomonas reinhardtii
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.1.3.1
C165S
behaves as the wild-type
Chlamydomonas reinhardtii
4.1.3.1
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
4.1.3.1
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
4.1.3.1
C178S
is totally inactive
Chlamydomonas reinhardtii
4.1.3.1
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
4.1.3.1
C301S
behaves as the wild-type
Chlamydomonas reinhardtii
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.1.3.1
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
4.1.3.1
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] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.1.3.1
46830
-
mutant C247S mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46860
-
mutant C178S mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46870
-
mutant C178S mass spectrometry, after 1 h of GSSG-treated ICL; wild-type, mass spectrometry
Chlamydomonas reinhardtii
4.1.3.1
46890
-
mutant C178S mass spectrometry, after 24 h of GSSG-treated ICL; mutant C178S mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47120
-
mutant C247S mass spectrometry, after 1 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47140
-
mutant C247S mass spectrometry, after 24 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47160
-
mutant C247S mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47170
-
wild-type, mass spectrometry, after 1 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47190
-
wild-type, mass spectrometry, after 5 h of GSSG-treated ICL
Chlamydomonas reinhardtii
4.1.3.1
47510
-
wild-type, mass spectrometry, after 24 h of GSSG-treated ICL
Chlamydomonas reinhardtii
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.1.3.1
to homogeneity, by Ni2+ affinity chromatography
Chlamydomonas reinhardtii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
4.1.3.1
9.5
-
purified ICL
Chlamydomonas reinhardtii
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
4.1.3.1
-20°C
Chlamydomonas reinhardtii
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.1.3.1
isocitrate
-
704605
Chlamydomonas reinhardtii
succinate + glyoxylate
-
-
-
?