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Literature summary for 4.1.3.1 extracted from

  • Bedhomme, M.; Zaffagnini, M.; Marchand, C.H.; Gao, X.H.; Moslonka-Lefebvre, M.; Michelet, L.; Decottignies, P.; Lemaire, S.D.
    Regulation by glutathionylation of isocitrate lyase from Chlamydomonas reinhardtii (2009), J. Biol. Chem., 284, 36282-36291.
    View publication on PubMedView publication on EuropePMC

Activating Compound

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(Commentary)

Cloned (Comment) Organism
wild-type and mutants cloned by the pET-3c-HIS/Escherichia coli BL21 expression system Chlamydomonas reinhardtii

Protein Variants

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

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 [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

Organism UniProt Comment Textmining
Chlamydomonas reinhardtii
-
-
-

Purification (Commentary)

Purification (Comment) Organism
to homogeneity, by Ni2+ affinity chromatography Chlamydomonas reinhardtii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
9.5
-
purified ICL Chlamydomonas reinhardtii

Storage Stability

Storage Stability Organism
-20°C Chlamydomonas reinhardtii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate
-
Chlamydomonas reinhardtii succinate + glyoxylate
-
?

Synonyms

Synonyms Comment Organism
ICL
-
Chlamydomonas reinhardtii
isocitrate lyase
-
Chlamydomonas reinhardtii