Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant Strep-tagged enzyme | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
purified enzyme, hanging drop vapour diffusion method, mixing of 0.0015 ml of 7 mg/mL CDO mutant C93G in 10 mM sodium phosphate and 20 mM NaCl, pH 7.5, and 0.0006 ml of crushed wild-type CDO crystal seeds in their own growth solution consisting of 25% w/v PEG 1500, 13 mM succinate, 44 mM sodium phosphate, and 44 mM glycine, pH 5.2, with 0.0015 ml of reservoir buffer containing 26% w/v PEG 4000, 200 mM ammonium acetate, and 100 mM sodium citrate, pH 6.3, equilibration against reservoir solution, X-ray diffraction structure determination and analysis at 1.82 A resolution | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
C93G | site-directed mutagenesis, structure comparison to the wild-type enzyme | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state Michaelis-Menten kinetics | Rattus norvegicus | |
0.002 | - |
L-cysteine | 20% cross-linked wild-type enzyme, pH 9.1, temperature not specified in the publication | Rattus norvegicus | |
0.003 | - |
L-cysteine | not cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus | |
0.005 | - |
L-cysteine | 60% cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | active site bound, reconstitution of the purified enzyme | Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cysteine + O2 | Rattus norvegicus | - |
3-sulfinoalanine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | P21816 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant Strep-tagged wild-type and mutant enzyme by affinity chromatography to over 95% purity | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cysteine + O2 | - |
Rattus norvegicus | 3-sulfinoalanine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CDO | - |
Rattus norvegicus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
L-cysteine | 20% cross-linked wild-type enzyme, pH 9.1, temperature not specified in the publication | Rattus norvegicus | |
0.25 | - |
L-cysteine | 60% cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus | |
0.33 | - |
L-cysteine | not cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
pH-dependence of wild-type and mutant enzyme kinetics, detailed overview | Rattus norvegicus |
7.7 | - |
60% cross-linked wild-type enzyme | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
Cys-Tyr cofactor | - |
Rattus norvegicus |
General Information | Comment | Organism |
---|---|---|
additional information | modeling the pH-dependence of cysteine dioxygenation | Rattus norvegicus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.056 | - |
L-cysteine | 20% cross-linked wild-type enzyme, pH 9.1, temperature not specified in the publication | Rattus norvegicus | |
0.056 | - |
L-cysteine | 60% cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus | |
0.095 | - |
L-cysteine | not cross-linked wild-type enzyme, pH 7.7, temperature not specified in the publication | Rattus norvegicus |