Cloning of CYP11B1 and CYP11B2 from normal human adrenal and their functional expression in COS-7 and V79 Chinese hamster cells.
The cDNAs of human 11 beta hydroxylase (CYP11B1) and aldosterone synthase (CYP11B2) were cloned from surgically removed normal human adrenal using polymerase chain reaction. The cloned cDNAs were transfected into COS-7 cells and steroid hydroxylase activity of the two cytochromes P450 expressed was determined in order to elucidate their the functional characteristics. Stable expression of human CYP11B1 or CYP11B2 was performed in V79 hamster cells and confirmed by Southern blotting, Northern blotting, and enzymatic activity. Interestingly, recombinant V79 cells were able to support CYP11B1 and CYP11B2 dependent steroid conversion without additional heterologous expression of the corresponding electron donor system.
Denner
K
K
Department of Chemistry, Freie Universität Berlin.
Doehmer
J
J
Bernhardt
R
R
D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003345:Corticosterone / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D019405:Cytochrome P-450 CYP11B2; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000235:genetics*; D003900:Desoxycorticosterone / Q000378:metabolism; D006801:Humans; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D012016:Reference Values; D013252:Steroid 11-beta-Hydroxylase / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D011994:Recombinant Proteins; D003900:Desoxycorticosterone; D003577:Cytochrome P-450 Enzyme System; D019405:Cytochrome P-450 CYP11B2; D013252:Steroid 11-beta-Hydroxylase; D003345:Corticosterone