Modulation of aldosterone biosynthesis by adrenodoxin mutants with different electron transport efficiencies.
Aldosterone biosynthesis is highly regulated on different levels by hormones, potassium, lipid composition of the membrane and the molecular structure of its gene. Here, the influence of the electron transport efficiency from adrenodoxin (Adx) to CYP11B1 on the activities of bovine CYP11B1 has been investigated using a liposomal reconstitution system with truncated mutants of Adx. It could be clearly demonstrated that Adx mutants Adx 4-114 and Adx 4-108, possessing enhanced electron transfer abilities, produce increases in corticosterone and aldosterone biosynthesis. Based on the Vmax values of corticosterone and aldosterone formation, Adx 4-108 and Adx 4-114 enhance corticosterone synthesis 1.3-fold and aldosterone formation threefold and twofold, respectively. The production of 18-hydroxycorticosterone was changed only slightly in these Adx mutants. The effect of Adx 1-108 on the product patterns of bovine CYP11B1, human CYP11B1 and human CYP11B2 was confirmed in COS-1 cells by cotransfection of CYP11B- and Adx-containing expression vectors. It could be shown that Adx 1-108 enhances the formation of aldosterone by bovine CYP11B1 and by human CYP11B2, and stimulates the production of corticosterone by bovine CYP11B1 and human CYP11B1 and CYP11B2 also.
Cao
P R
PR
Universität des Saarlandes, Fachrichtung 12.4-Biochemie, Saarbrücken, germany.
Bernhardt
R
R
D000325:Adrenodoxin / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D000450:Aldosterone / Q000096:biosynthesis*; D000818:Animals; D002417:Cattle; D003345:Corticosterone / Q000096:biosynthesis; D019405:Cytochrome P-450 CYP11B2 / Q000378:metabolism; D003900:Desoxycorticosterone / Q000378:metabolism; D004579:Electron Transport; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism; D009154:Mutation; D010446:Peptide Fragments / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D013252:Steroid 11-beta-Hydroxylase / Q000187:drug effects / Q000378:metabolism*
D008081:Liposomes; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000325:Adrenodoxin; D003900:Desoxycorticosterone; D000450:Aldosterone; D019405:Cytochrome P-450 CYP11B2; D013252:Steroid 11-beta-Hydroxylase; D005287:Ferredoxin-NADP Reductase; D003345:Corticosterone