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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
L-cysteine + O2 = 3-sulfinoalanine
reaction mechanism with internal electron transfer involving the ferric/ferrous enzyme forms, formation of a transient substrate-bound FeIII-superoxo species, overview
concentration of FeIII-CDO is highly variable and often does not exceed 5% relative to the catalytically active ferrous enzyme, ferric enzyme is catalytically inactive
a a mononuclear nonheme iron(II)-dependent enzyme. It possesses both an unusual 3-His facial ligation sphere to the iron center and a rare Cys-Tyr crosslink near the active site
presence of 0.1 mol Fe2+, absence of EDTA. Cysteine sulfinic acid content is determined by ion-paired reverse-phase chromatopraphy using UV detection at 215 nm.
presence of 0.1 mol Fe2+, absence of EDTA. Cysteine sulfinic acid content is determined by ion-paired reverse-phase chromatopraphy using UV detection at 215 nm.
lack of methionine sulfoxide reductase MsrA in liver of MsrA -/- mice leads to a significant drop in the cellular level of thiol groups and lowers the level of cysteine dioxygenase expression. Following selenium deficient diet applied to decrease the expression levels of selenoproteins like MsrB, the latter effect is maintained while the basal levels of thiol decreased in both wild-type strains and strains deficient for methionine dioxygenase
CDO knockout female mice exhibit severe defects in mammary branching morphogenesis and ductal elongation, resulting in poor lactation. CDO contributes to the luminal epithelial cell differentiation, proliferation, and apoptosis mainly through its downstream product cysteine sulfinic acid. Exogenous supplementation of cysteine sulfinic acid rescues the defects in CDO knockout mice and enhances ductal growth in wild-type mice
CDO-/- mouse sperm demonstrates a severe lack of in vitro fertilization ability. Acrosome exocytosis and tyrosine phosphorylation profiles in response to stimuli are normal. CDO-/- sperm has a slight increase in head abnormalities. Taurine and hypotaurine concentrations in CDO-/- sperm decrease in the epididymal intraluminal fluid and sperm cytosol. No evidence of antioxidant protection against lipid peroxidation is found. CDO-/- sperm exhibits severe defects in volume regulation, swelling in response to the relatively hypo-osmotic conditions found in the female reproductive tract
transcription factor NRF2, i.e. nuclear factor-erythroid 2 p45-related factor two, promotes the accumulation of intracellular cysteine and engages the cysteine homeostatic control mechanism mediated by cysteine dioxygenase 1
cysteine dioxygenase is a mononuclear nonheme iron(II)-dependent enzyme critical for maintaining appropriate cysteine and taurine levels in eukaryotic systems
NO as a substrate analogue for O2 is used to prepare a species that mimics the geometric and electronic structures of an early reaction intermediate, analysis by magnetic circular dichroism, electron paramagnetic resonance, and electronic absorption spectroscopies as well as computational methods including density functional theory and semiempirical calculations, quantum mechanics/molecular mechanics calculations, overview. The NO adducts of Cys- and selenocysteine (Sec)-bound Fe(II)CDO exhibit virtually identical electronic properties
the catalytic cycle of CDO can be primed by one electron through chemical oxidation to produce CDO with ferric iron in the active site. The C93-Y157 pair of mammalian CDO is catalytically essential. The monoanionic active site contains a 5- or 6-coordinate ferrous iron with solvent molecules serving as the non-protein ligands. In the absence of substrate and/or cofactor, the reduced active site is unreactive toward O2
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystals are grown at 25°C by hanging-drop vapor-diffusion. The reservoir contains 20% methosylpolyethylene glycol 5000, 160 mM CaCl2, and 100 mM 2-morpholinjoethane-sulfonic acid (pH 6.5). Hanging drops consist of 2 microl of protein solution mixed with 2 microl of reservoir solution. Structure is solved to a nominal 1.75 A resolution.
electron paramagnetic study of substrate-O2 binding. Ordered binding of L-cysteine prior to NO and presumably O2. Upon addition of NO to cysteine dioxygenase in the presence of substrate L-cysteine, a low-spin(FeNO)7 signal with spin S of 1/2that accounts for 85% of the iron within the enzyme develops. Substitution of L-cysteine with isosteric substrate analogues cysteamine, 3-mercaptopropionic acid, and propane thiol does not produce any analogous signals.The unusual (FeNO)7, spin 1/2 electronic configuration adopted by the substrate-bound iron-nitrosyl cysteine dioxygenase is a result of the bidentate thiol/amine coordination of L-cysteine in the NO-bound active site
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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
the fusion protein is purified by immobilized metal (Ni2+) affinity chromatography. The liberated enzyme is separated from the His-8-maltose binding protein by immobilized metal affinity chromatography, further purified by gel filtration chromatography, and concentrated. EDTA is obmitted from the buffers used to purifiy enzyme for catalytic studies, and the gel filtration is not used.
recombinant N-terminal maltose-binding protein fusion enzyme from Escherichia coli strain BL21(DE3) by anion exchange and amylose affinity chromatography
recombinant N-terminal maltose-binding protein fusion enzyme from Escherichia coli strain BL21(DE3)RIPL by anion exchange and amylose affinity chromatography
cloning of the gene and comparison of the gene to the known rat and human genes, sequencing, profiling of the enzyme mRNA and protein levels in mouse tissues
lack of methionine sulfoxide reductase MsrA in liver of MsrA -/- mice leads to a significant drop in the cellular level of thiol groups and lowers the level of cysteine dioxygenase expression. Following selenium deficient diet applied to decrease the expression levels of selenoproteins like MsrB, the latter effect is maintained while the basal levels of thiol decreased in both wild-type strains and strains deficient for methionine dioxygenase
Characterization of the nitrosyl adduct of substrate-bound mouse cysteine dioxygenase by electron paramagnetic resonance: electronic structure of the active site and mechanistic implications
Spectroscopic and computational characterization of substrate-bound mouse cysteine dioxygenase: nature of the ferrous and ferric cysteine adducts and mechanistic implications
Single turnover of substrate-bound ferric cysteine dioxygenase with superoxide anion: enzymatic reactivation, product formation, and a transient intermediate
Spectroscopic and computational characterization of the NO adduct of substrate-bound Fe(II) cysteine dioxygenase: insights into the mechanism of O2 activation