Cloning and expression of rat 25-hydroxyvitamin D3-1α-hydroxylase cDNA

T. Shinki, H. Shimada, Shu Wakino, H. Anazawa, M. Hayashi, T. Saruta, H. F. DeLuca, T. Suda

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

A full-length cDNA for the rat kidney mitochondrial cytochrome P450 mixed function oxidase, 25-hydroxyvitamin D3-1α-hydroxylase (P4501α), was cloned from a vitamin D-deficient rat kidney cDNA library and subcloned into the mammalian expression vector pcDNA 3.1(+). When P4501α cDNA was transfected into COS-7 transformed monkey kidney cells, they expressed 25- hydroxyvitamin D3-1α-hydroxylase activity. The sequence analysis showed that P4501α was of 2,469 bp long and contained an ORF encoding 501 amino acids. The deduced amino acid sequence showed a 53% similarity and 44% identity to the vitamin D3-25-hydroxylase (CYP27), whereas it has 42.6% similarity and 34% identity with the 25-hydroxyvitamin D3-24-hydroxylase (CYP24). Thus, it composes a new subfamily of the CYP27 family. Further, it is more closely related to the CYP27 than to the CYP24. The expression of P4501α mRNA was greatly increased in the kidney of vitamin D-deficient rats. In rats with the enhanced renal production of 1α,25-dihydroxyvitamin D3 (rats fed a low Ca diet), P4501α mRNA was greatly increased in the renal proximal convoluted tubules.

Original languageEnglish
Pages (from-to)12920-12925
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number24
DOIs
Publication statusPublished - 1997

Fingerprint

Calcifediol
Mixed Function Oxygenases
Organism Cloning
Complementary DNA
Kidney
Vitamin D
Cholestanetriol 26-Monooxygenase
Proximal Kidney Tubule
Messenger RNA
Calcitriol
Gene Library
Cytochrome P-450 Enzyme System
Open Reading Frames
Haplorhini
Sequence Analysis
Amino Acid Sequence
Diet
Amino Acids

Keywords

  • 1α-hydroxylase
  • 24-hydroxylase
  • P450
  • Vitamin D metabolism

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Cloning and expression of rat 25-hydroxyvitamin D3-1α-hydroxylase cDNA. / Shinki, T.; Shimada, H.; Wakino, Shu; Anazawa, H.; Hayashi, M.; Saruta, T.; DeLuca, H. F.; Suda, T.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, No. 24, 1997, p. 12920-12925.

Research output: Contribution to journalArticle

Shinki, T. ; Shimada, H. ; Wakino, Shu ; Anazawa, H. ; Hayashi, M. ; Saruta, T. ; DeLuca, H. F. ; Suda, T. / Cloning and expression of rat 25-hydroxyvitamin D3-1α-hydroxylase cDNA. In: Proceedings of the National Academy of Sciences of the United States of America. 1997 ; Vol. 94, No. 24. pp. 12920-12925.
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