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Literature summary for 1.14.15.18 extracted from
- Metabolism of substrates incorporated into phospholipid vesicles by mouse 25-hydroxyvitamin D3 1alpha-hydroxylase (CYP27B1) (2010), J. Steroid Biochem. Mol. Biol., 119, 171-179.
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| adrenodoxin | mouse and bovine adrenodoxin display similar abilities to support CYP27B1 activity | Mus musculus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression of His-tagged CYP27B1 in Escherichia coli strain JM109 | Mus musculus |
General Stability
| General Stability | Organism |
|---|---|
| CYP27B1 is labile during purification despite the presence of 20% glycerol as a stabilizing agent | Mus musculus |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 25-hydroxyvitamin D2 | CYP27B1 shows inhibition when substrate concentrations in the membrane are greater than 4 times Km, more pronounced with 25-hydroxyvitamin D3 than with 25-hydroxyvitamin D2 | Mus musculus |  |
| 25-hydroxyvitamin D3 | CYP27B1 shows inhibition when substrate concentrations in the membrane are greater than 4 times Km, more pronounced with 25-hydroxyvitamin D3 than with 25-hydroxyvitamin D2 | Mus musculus | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics, and by a kinetic model with substrate binding at two inhibitory sites, overview | Mus musculus | |
| 4.6 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 4.8 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 5.9 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 9.7 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 26 | - |
(24R),25-dihydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 49 | - |
20-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 25-hydroxyvitamin D2 + NADPH + H+ + O2 | Mus musculus | - |
1,25-dihydroxyvitamin D2 + NADP+ + H2O | - |
? | |
| 25-hydroxyvitamin D3 + NADPH + H+ + O2 | Mus musculus | - |
1,25-dihydroxyvitamin D3 + NADP+ + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mus musculus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged CYP27B1 from Escherichia coli strain JM109 by nickel affinity chromatoghraphy, followed by affinity chromatography on a bovine adrenodoxin-coupled resin with elution by 1.0% CHAPS | Mus musculus |
Storage Stability
| Storage Stability | Organism |
|---|---|
| 4°C, prolonged standing of the enzyme, including overnight dialysis, causes marked loss of the holo-enzyme | Mus musculus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (24R),25-dihydroxyvitamin D3 + NADPH + O2 | - |
Mus musculus | 1alpha,24,25-trihydroxyvitamin D3 + NADP+ + H2O | - |
? | |
| 20-hydroxyvitamin D3 + NADPH + O2 | - |
Mus musculus | 1alpha,20-dihydroxyvitamin D3 + NADP+ + H2O | - |
? | |
| 25-hydroxyvitamin D2 + NADPH + H+ + O2 | - |
Mus musculus | 1,25-dihydroxyvitamin D2 + NADP+ + H2O | - |
? | |
| 25-hydroxyvitamin D3 + NADPH + H+ + O2 | - |
Mus musculus | 1,25-dihydroxyvitamin D3 + NADP+ + H2O | - |
? | |
| additional information | 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in vesicles undergo 1alpha-hydroxylation with similar kinetics. CYP27B1 also catalyzes 1alpha-hydroxylation of vesicle-associated 24R,25-dihydroxyvitamin D3 and 20-hydroxyvitamin D3, and 25-hydroxylation of 1alpha-hydroxyvitamin D3 and 1alpha-hydroxyvitamin D2, but with much lower efficiency than for 25(OH)D3, reactions in reconstituted phospholipid vesicles with the substrates associating to the membrane, effect of vesicle phospholipid composition on metabolism of 25(OH)D3 by CYP27B1, overview | Mus musculus | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 25-hydroxyvitamin D3 1alpha-hydroxylase | - |
Mus musculus |
| CYP27B1 | - |
Mus musculus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Mus musculus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.14 | - |
20-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 0.68 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.8 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.82 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 0.9 | - |
(24R),25-dihydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.92 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.4 | - |
assay at | Mus musculus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | - |
Mus musculus | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.003 | - |
20-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 0.035 | - |
(24R),25-dihydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.094 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
| 0.115 | - |
25-hydroxyvitamin D3 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.17 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, Michaelis-Menten kinetic parameters | Mus musculus | |
| 0.171 | - |
25-hydroxyvitamin D2 | pH 7.4, 37°C, parameters using a kinetic model with substrate binding at two inhibitory sites | Mus musculus | |
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