Structural analysis of cloned cDNA for mRNA of microsomal cytochrome P-450(C21) which catalyzes steroid 21-hydroxylation in bovine adrenal cortex

H. Yoshioka, Ken-Ichirou Morohashi, K. Sogawa, M. Yamane, S. Kominami, S. Takemori, Y. Okada, T. Omura, Y. Fujii-Kuriyama

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Abstract

We have isolated cDNA clones of the mRNA for cytochrome P-450 that catalzyes the steroid C-21 hydroxylation (P-450(C21)), which specifically catalyzes 21-hydroxylation of steroids in the microsomes of bovine adrenal cortex by using synthetic oligonucleotides as probes. Sequence determination of the cloned cDNA showed that it contains 2157 nucleotides and a poly(A) chain and that a single open reading frame of 1488 nucleotides codes for a polypeptide of 496 amino acids with a molecular weight of 50,113. The deduced amino acid composition is agreement with that determined by direct amino acid analysis of purified P-450(C21) and the predicted primary structure contained amino acid sequences of N-terminal region and two internal tryptic fragments of the protein so far analyzed. Comparing the amino acid sequence with those of other forms of P-450 reveals that a conserved amino acid sequence containing a putative heme-binding cysteine is present in the equivalent position, proximate to the COOH terminus of the molecules and that P-450(C21) is phylogenically situated in an intermediate position between steroidogenic mitochondrial cytochrome P-450 which catalyzes the side-chain cleavage of cholesterol (P450(SCC)) and drug-metabolizing microsomal P-450s. However, the amino acid sequence of P-450(C21) is much close to that of drug-metabolizing P-450s than to that of P-450(SCC).

Original languageEnglish
Pages (from-to)4106-4109
Number of pages4
JournalJournal of Biological Chemistry
Volume261
Issue number9
Publication statusPublished - Dec 1 1986
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Yoshioka, H., Morohashi, K-I., Sogawa, K., Yamane, M., Kominami, S., Takemori, S., Okada, Y., Omura, T., & Fujii-Kuriyama, Y. (1986). Structural analysis of cloned cDNA for mRNA of microsomal cytochrome P-450(C21) which catalyzes steroid 21-hydroxylation in bovine adrenal cortex. Journal of Biological Chemistry, 261(9), 4106-4109.