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

doi:10.1093/oxfordjournals.jbchem.a123099

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

Biology of Sex Difference

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65 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 language	English
Pages (from-to)	635-640
Number of pages	6
Journal	Journal of biochemistry
Volume	107
Issue number	4
DOIs	https://doi.org/10.1093/oxfordjournals.jbchem.a123099
Publication status	Published - Apr 1990

All Science Journal Classification (ASJC) codes

Biochemistry
Molecular Biology

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

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.",

author = "Morohashi, {Ken Ichirou} and Yasuki Nonaka and Shirou Kirita and Osamu Hatano and Akira Takakusu and Mitsuhiro Okamoto and Tsuneo Omura",

year = "1990",

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doi = "10.1093/oxfordjournals.jbchem.a123099",

language = "English",

volume = "107",

pages = "635--640",

journal = "Journal of Biochemistry",

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T1 - Enzymatic activities of P-450(llβ)S expressed by two cDNAs in COS-7 cells

AU - Morohashi, Ken Ichirou

AU - Nonaka, Yasuki

AU - Kirita, Shirou

AU - Hatano, Osamu

AU - Takakusu, Akira

AU - Okamoto, Mitsuhiro

AU - Omura, Tsuneo

PY - 1990/4

Y1 - 1990/4

N2 - 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.

AB - 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.

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JO - Journal of Biochemistry

JF - Journal of Biochemistry

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ER -

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

Abstract

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