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Literature summary extracted from

  • Sugimoto, H.; Shinkyo, R.; Hayashi, K.; Yoneda, S.; Yamada, M.; Kamakura, M.; Ikushiro, S.; Shiro, Y.; Sakaki, T.
    Crystal structure of CYP105A1 (P450SU-1) in complex with 1alpha,25-dihydroxyvitamin D3 (2008), Biochemistry, 47, 4017-4027.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.22 expression in Escherichia coli Streptomyces griseolus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.15.22 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

EC Number Protein Variants Comment Organism
1.14.15.22 I243A complete loss of activity Streptomyces griseolus
1.14.15.22 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
1.14.15.22 L180A complete loss of activity Streptomyces griseolus
1.14.15.22 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
1.14.15.22 R193K complete loss of activity Streptomyces griseolus
1.14.15.22 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
1.14.15.22 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
1.14.15.22 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
1.14.15.22 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
1.14.15.22 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
1.14.15.22 V181A complete loss of activity Streptomyces griseolus
1.14.15.22 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

EC Number Organism UniProt Comment Textmining
1.14.15.22 Streptomyces griseolus P18326
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Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.15.22 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 and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.22 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
-
 ?
1.14.15.22 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
-
 ?
1.14.15.22 vitamin D3 + O2 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
 Streptomyces griseolus 1alpha-hydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?
1.14.15.22 vitamin D3 + O2 + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+
-
 Streptomyces griseolus 25-hydroxyvitamin D3 + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O
-
 ?