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.
|Number of pages||6|
|Journal||Journal of biochemistry|
|Publication status||Published - Apr 1990|
All Science Journal Classification (ASJC) codes
- Molecular Biology