Purification of cytochrome P-450 catalyzing 25-hydroxylation of vitamin D3 from rat liver microsomes

Shin ichi Hayashi, Mitsuhide Noshiro, Kyuichiro Okuda

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)


Cytochrome P-450 catalyzing 25-hydroxylation of cholecalciferol (cytochrome P-450cc25) was purified from rat liver microsomes based on its catalytic activity at each purification step. The specific cytochrome P-450 content of the final preparation was 15.1 nmol/mg of protein. Reconstituted activity of 25-hydroxylation of cholecalciferol with the purified enzyme was 2.3 nmol/min/mg of protein, which was 4,300 times as high as that in microsomes. The minimum molecular weight of the enzyme was 50,000 based on SDS-polyacrylamide gel electrophoretogram. Amino terminal sequence of the P-450cc25 was H2N-Met-Asp-Pro-Val-Leu-Val-. Immunochemical study showed that the purified P-450cc25 was homogeneous and the cytochrome was immunochemically different from either cytochrome P-450(PB-1) or cytochrome P-448(MC-1).

Original languageEnglish
Pages (from-to)994-1000
Number of pages7
JournalBiochemical and biophysical research communications
Issue number3
Publication statusPublished - 1984 Jun 29
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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