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Literature summary for 1.14.15.16 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)

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

Protein Variants

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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
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

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Rattus norvegicus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
?
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
-
?
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
-
?
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
-
?
additional information human CYP24A1 catalyzes both C-23 and C-24 oxidation pathways of 1alpha,25-dihydroxyvitamin D3 Homo sapiens ?
-
?

Synonyms

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
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
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
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
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
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
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
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
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
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

Cofactor Comment Organism Structure
NADPH
-
Homo sapiens
NADPH
-
Rattus norvegicus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
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
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
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
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
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
20
-
1alpha,25-dihydroxyvitamin D3 mutant enzyme T416A, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Rattus norvegicus
20
-
1alpha,25-dihydroxyvitamin D3 wild type enzyme, in 100 mM Tris-HCl, pH 7.4, temperature not specified in the publication Homo sapiens
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
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