Cloned (Comment) | Organism |
---|---|
cloned in plasmid pVP16 to produce the enzyme in Escherichia coli B834 pRARE2, an N-terminal fusion to a His-8-maltose binding protein | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
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. | Mus musculus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | higher kcat and lower Km value, when the enzyme is expressed in medium containing extra iron and purified in buffers lacking EDTA. | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.1 | - |
L-cysteine | 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. | Mus musculus | |
3.4 | - |
L-cysteine | 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. | Mus musculus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | higher kcat and lower Km value, when the enzyme is expressed in medium containing extra iron and purified in buffers lacking EDTA. | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | P60334 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
additional information | the residues Tyr-157 and Cys-93 appear to be covalently linked between Tyr-157 CE and Cys-93 SG, because these two atoms lie within 2.2 A | Mus musculus |
Purification (Comment) | Organism |
---|---|
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. | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-cysteine + O2 | - |
Mus musculus | 3-sulfino-L-alanine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CDO | - |
Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.8 | - |
L-cysteine | presence of 0.1 mol Fe2+, absence of EDTA | Mus musculus | |
3.6 | - |
L-cysteine | presence of 0.1 mol Fe2+, absence of EDTA | Mus musculus |