Cloned (Comment) | Organism |
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expression of wild-type Vdh and mutant Vdh-K1 in Escherichia coli | Pseudonocardia autotrophica |
Crystallization (Comment) | Organism |
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purified recombinant wild-type Vdh and mutant Vdh-K1 in complex with substrates VD3 and 25(OH)VD3, 20 mg/ml wild-type protein in 20 mM Tris-HCl, pH 7.5, supplemented with 0-0.2% PMbetaCD for VD3 complex or 0-0.2% gammaCD for 25(OH)VD3 complex, 20 mg/ml mutant protein in 20 mM Tris-HCl, pH 7.5, supplemented with 20 mM NaCl, sitting-drop method, 20°C. Vdh-WT crystals are grown using reservoir solution containing 0.1 M BisTris, pH 7.5, 50 mM CaCl2, 40-120 mM NaCl or KCl, and 32-40% PEG 400 or PEG 550 monomethyl ether or 20% PEG 1000, Vdh-K1 is crystallized using reservoir solution containing 0.1 M calcium acetate and 10-14% PEG 3350, X-ray diffraction structure determination and analysis at 1.75-3.05 A resolution | Pseudonocardia autotrophica |
Protein Variants | Comment | Organism |
---|---|---|
T70R/V156L/E216M/E384R | mutant Vdh-K1, 22fold more active in the hydroxylation of VD3 to 25(OH)VD3 than wild-type Vdh in in vivo bioconversion using Escherichia coli cells. backbone and side chain conformations at the active-site pocket are identical among the substrate-free, VD3-bound, and 25(OH)VD3-bound forms of Vdh-K1, and no induced-fit mechanisms are observed, although VD3 is asymmetric | Pseudonocardia autotrophica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Pseudonocardia autotrophica | Vdh, a cytochrome P450 enzyme, is responsible for the biocatalytic conversion of vitamin D3, catalyzing the two-step hydroxylation of VD3, i.e. the conversion of VD3 to 25-hydroxyvitamin D3 (25(OH)VD3) and then of 25(OH)VD3 to 1alpha,25(OH)2VD3, a hormonal form of VD3. Anti-parallel substrate binding modes enable sequential hydroxylation, structure-function analysis, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudonocardia autotrophica | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type Vdh and mutant Vdh-K1 from Escherichia coli | Pseudonocardia autotrophica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Vdh, a cytochrome P450 enzyme, is responsible for the biocatalytic conversion of vitamin D3, catalyzing the two-step hydroxylation of VD3, i.e. the conversion of VD3 to 25-hydroxyvitamin D3 (25(OH)VD3) and then of 25(OH)VD3 to 1alpha,25(OH)2VD3, a hormonal form of VD3. Anti-parallel substrate binding modes enable sequential hydroxylation, structure-function analysis, overview | Pseudonocardia autotrophica | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | Vdh structure analysis, overview | Pseudonocardia autotrophica |
Synonyms | Comment | Organism |
---|---|---|
25(OH)VD3 1alpha-hydroxylase | - |
Pseudonocardia autotrophica |
VD3 25-hydroxylase | - |
Pseudonocardia autotrophica |
VDH | - |
Pseudonocardia autotrophica |
vitamin D3 hydroxylase | - |
Pseudonocardia autotrophica |