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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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 - Jan 1 1990

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Desoxycorticosterone
COS Cells
Corticosterone
Microscopic examination
18-Hydroxycorticosterone
Complementary DNA
Hydroxylation
Mitochondria
Electroporation
Immunoelectron Microscopy
Aldosterone
Fluorescence Microscopy
Immunoblotting
Plasmids
Clone Cells
Substrates
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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Enzymatic activities of P-450(llβ)S expressed by two cDNAs in COS-7 cells. / Morohashi, Ken-Ichirou; Nonaka, Yasuki; Kirita, Shirou; Hatano, Osamu; Takakusu, Akira; Okamoto, Mitsuhiro; Omura, Tsuneo.

In: Journal of Biochemistry, Vol. 107, No. 4, 01.01.1990, p. 635-640.

Research output: Contribution to journalArticle

Morohashi, K-I, Nonaka, Y, Kirita, S, Hatano, O, Takakusu, A, Okamoto, M & Omura, T 1990, 'Enzymatic activities of P-450(llβ)S expressed by two cDNAs in COS-7 cells', Journal of Biochemistry, vol. 107, no. 4, pp. 635-640. https://doi.org/10.1093/oxfordjournals.jbchem.a123099
Morohashi, Ken-Ichirou ; Nonaka, Yasuki ; Kirita, Shirou ; Hatano, Osamu ; Takakusu, Akira ; Okamoto, Mitsuhiro ; Omura, Tsuneo. / Enzymatic activities of P-450(llβ)S expressed by two cDNAs in COS-7 cells. In: Journal of Biochemistry. 1990 ; Vol. 107, No. 4. pp. 635-640.
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AU - Takakusu, Akira

AU - Okamoto, Mitsuhiro

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