Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Streptomyces griseolus |
Crystallization (Comment) | Organism |
---|---|
wild-type to 1.5 A resolution, and mutant R84A, native protein and in complex with 1alpha,25-dihydroxyvitamin D3. The compound is positioned 11 A from the iron atom along the I helix within the pocket. The loss of two hydrogen bonds in the R84A mutant increases the adaptability of the B and F helices, creating a transient binding site | Streptomyces griseolus |
Protein Variants | Comment | Organism |
---|---|---|
I243A | complete loss of activity | Streptomyces griseolus |
I293A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 5.4 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 3.4 mmol of product per min and mol of protein | Streptomyces griseolus |
L180A | complete loss of activity | Streptomyces griseolus |
R193A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 0.48 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 0.72 mmol of product per min and mol of protein | Streptomyces griseolus |
R193K | complete loss of activity | Streptomyces griseolus |
R193Q | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 0.33 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 0.31 mmol of product per min and mol of protein | Streptomyces griseolus |
R73A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 30.9 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 20.8 mmol of product per min and mol of protein | Streptomyces griseolus |
R84A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 75.7 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 34.5 mmol of product per min and mol of protein | Streptomyces griseolus |
R89A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 0.25 mmol of product per min and mol of protein, no 25-hydroxyvitamin D3 hydroxylation activity at C1alpha | Streptomyces griseolus |
S236A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 4.6 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 3.4 mmol of product per min and mol of protein | Streptomyces griseolus |
V181A | complete loss of activity | Streptomyces griseolus |
V88A | 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 1.52 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 0.34 mmol of product per min and mol of protein | Streptomyces griseolus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces griseolus | P18326 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
wild-type, 1alpha-hydroxyvitamin D3 hydroxylation activity at C25 2.8 mmol of product per min and mol of protein, 25-hydroxyvitamin D3 hydroxylation activity at C1alpha 2.1 mmol of product per min and mol of protein | Streptomyces griseolus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1alpha-hydroxyvitamin D3 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Streptomyces griseolus | 1alpha,25-dihydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
25-hydroxyvitamin D3 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ + O2 | - |
Streptomyces griseolus | 1alpha,25-dihydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
vitamin D3 + O2 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | - |
Streptomyces griseolus | 1alpha-hydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? | |
vitamin D3 + O2 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | - |
Streptomyces griseolus | 25-hydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O | - |
? |