Enzymatic activities of P-450(llβ)S expressed by two cDNAs in COS-7 cells

Ken Ichirou Morohashi, Yasuki Nonaka, Shirou Kirita, Osamu Hatano, Akira Takakusu, Mitsuhiro Okamoto, Tsuneo Omura

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64 Citations (Scopus)

Abstract

Expression plasmids were constructed using two cDNA clones of P-450(llβ), pcP-450-(llβ)-2, and pcP-450(llβ)-3 (Morohashi et al. (1987) J. Biochenu 102, 559-568 and Kirita et al. (1988) J. Biochem. 104, 683-686), and introduced into COS-7 cells by electroporation. The expression of P-450(llβ) proteins and their localization in the mitochondria were demonstrated by immunoblotting, immunofluorescence microscopy, and immunoelectron microscopy. The enzymatic activities of the expressed P-450(llβ)s were determined using deoxycorticosterone (DOC), deoxycortisol, and corticosterone as substrates. Though the activities of the two P-450(llβ)s for 11-, 18-, and 19-hydroxylation of DOC were almost equal, the production of 18-hydroxycorticosterone and aldosterone from corticosterone by P-450(llβ)-3 was greater than that by P-450(llβ)-2.

Original languageEnglish
Pages (from-to)635-640
Number of pages6
JournalJournal of biochemistry
Volume107
Issue number4
DOIs
Publication statusPublished - Apr 1990

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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