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Literature summary extracted from

  • Hamamoto, H.; Kusudo, T.; Urushino, N.; Masuno, H.; Yamamoto, K.; Yamada, S.; Kamakura, M.; Ohta, M.; Inouye, K.; Sakaki, T.
    Structure-function analysis of vitamin D 24-hydroxylase (CYP24A1) by site-directed mutagenesis: amino acid residues responsible for species-based difference of CYP24A1 between humans and rats (2006), Mol. Pharmacol., 70, 120-128.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.15.16 mutant enzymes are expressed in Escherichia coli BL21(DE3) cells Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
1.14.15.16 I500F the mutant shows quite a different metabolism of 1alpha,25-dihydroxyvitamin D3 from both human and rat CYP24A1 Rattus norvegicus
1.14.15.16 I500L the mutant also shows the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 I500T the mutant also shows the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 I500V the mutant also shows the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 T416A the metabolic pattern by the mutant remains rat CYP24A1wild type Rattus norvegicus
1.14.15.16 T416F the mutant has reaction specificity similar to human CYP24A1 by also showing the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 T416I the mutant has reaction specificity similar to human CYP24A1 by also showing the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 T416M the mutant also shows the C-23 oxidation pathway Rattus norvegicus
1.14.15.16 T416S the metabolic pattern by the mutant remains rat CYP24A1wild type Rattus norvegicus
1.14.15.16 T416V the mutant has reaction specificity similar to human CYP24A1 by also showing the C-23 oxidation pathway Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.15.16 0.00014
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500T, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00019
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00022
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416S, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00024
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416A, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.0003
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500L, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00035
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Homo sapiens
1.14.15.16 0.00066
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416M, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00104
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416V, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.00114
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500F, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus

Organism

EC Number Organism UniProt Comment Textmining
1.14.15.16 Homo sapiens
-
-
-
1.14.15.16 Rattus norvegicus
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.15.16 1alpha,25-dihydroxyvitamin D3 + NADPH + H+ + O2 the C-23 oxidation pathway Homo sapiens 1alpha,23S,25,26-tetrahydroxyvitamin D3 + 1alpha,23S,25-trihydroxyvitamin D3 + 25,26,27-trinor-23-ene-1alpha-hydroxyvitamin D3 + NADP+ + H2O
-
 ?
1.14.15.16 1alpha,25-dihydroxyvitamin D3 + NADPH + H+ + O2 the C-23 oxidation pathway, which is almost not shown by rat CYP24A1 Rattus norvegicus 1alpha,23S,25,26-tetrahydroxyvitamin D3 + 1alpha,23S,25-trihydroxyvitamin D3 + 25,26,27-trinor-23-ene-1alpha-hydroxyvitamin D3 + NADP+ + H2O
-
 ?
1.14.15.16 1alpha,25-dihydroxyvitamin D3 + NADPH + H+ + O2 the C-24 oxidation pathway Homo sapiens 24-oxo-1,23,25-trihydroxyvitamin D3 + 24,25,26,27-tetranor-1,23-dihydroxyvitamin D3 + 1alpha,24R,25-trihydroxyvitamin D3 + 24-oxo-1,25-dihydroxyvitamin D3 + NADP+ + H2O
-
 ?
1.14.15.16 1alpha,25-dihydroxyvitamin D3 + NADPH + H+ + O2 the C-24 oxidation pathway Rattus norvegicus 24-oxo-1,23,25-trihydroxyvitamin D3 + 24,25,26,27-tetranor-1,23-dihydroxyvitamin D3 + 1alpha,24R,25-trihydroxyvitamin D3 + 24-oxo-1,25-dihydroxyvitamin D3 + NADP+ + H2O
-
 ?
1.14.15.16 additional information human CYP24A1 catalyzes both C-23 and C-24 oxidation pathways of 1alpha,25-dihydroxyvitamin D3 Homo sapiens ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.15.16 CYP24A1
-
 Homo sapiens
1.14.15.16 CYP24A1
-
 Rattus norvegicus
1.14.15.16 vitamin D 24-hydroxylase
-
 Homo sapiens
1.14.15.16 vitamin D 24-hydroxylase
-
 Rattus norvegicus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.15.16 0.0008
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500L, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.002
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500F, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.002
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500T, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.0035
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416M, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.004
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416A, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.004
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.005
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416S, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 0.007
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Homo sapiens
1.14.15.16 0.012
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416V, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.15.16 NADPH
-
 Homo sapiens
1.14.15.16 NADPH
-
 Rattus norvegicus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.15.16 1.7
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500F, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 2.8
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500L, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 5.3
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416M, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 11.8
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416V, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 15.3
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme I500T, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 20
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416A, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 20
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Homo sapiens
1.14.15.16 23.3
-
 1alpha,25-dihydroxyvitamin D3 mutant enzyme T416S, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
1.14.15.16 23.3
-
 1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus