Differential expression of cytochrome P-450 1 and related forms in rabbit liver and kidney.
Monoclonal antibodies developed to cytochrome P-450 1, some of which react with proteins in addition to P-450 1, were used to investigate the differential expression of P-450 1 dependent 21-hydroxylase activity in renal tissue of rabbits exhibiting differences in hepatic 21-hydroxylase activity. Using immunohistochemical techniques, the monoclonal antibodies, 2F5 and 3C3, localized protein in the S2 and S3 segments of the proximal tubule in the renal cortex. These two monoclonal antibodies, 2F5 and 3C3, reacted with a kidney protein that migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a relative electrophoretic mobility that did not correspond to known rabbit hepatic isozymes and was termed P-450 K. Antibodies specific for P-450 1 and 3b, 1F11 and 8-27, respectively, produced no staining in kidney. The protein recognized by the 2F5 and 3C3 antibodies is immunologically distinct from cytochrome P-450s 1, 2, and 3b. The rate of 21-hydroxylation of progesterone was shown to be approximately 100-fold less in kidney than liver microsomes where this pathway is largely catalyzed by P-450 1. The activity of the kidney microsomes was not inhibited by antibodies directed to P-450 1. In addition, the variation observed for the 21-hydroxylase activity in the hepatic microsomal fraction of outbred New Zealand white rabbits was not evident in kidney microsomes from these same animals. The 2F5 antibody was found, however, to be inhibitory (about 50%) of the 11-hydroxylation of lauric acid in kidney microsomes. This suggests that P-450 K participates in lauric acid 11-hydroxylase activity. The treatment of rabbits with phenobarbital, but not 2,3,7,8-tetrachlorodibenzo-p-dioxin, was found to induce the levels of P-450 K.
D000818:Animals; D000911:Antibodies, Monoclonal; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006651:Histocytochemistry; D007159:Immunologic Tests; D007527:Isoenzymes / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology*; D007687:Kidney Tubules, Proximal / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D011374:Progesterone / Q000378:metabolism; D011817:Rabbits; D013255:Steroid 21-Hydroxylase / Q000378:metabolism*; D013250:Steroid Hydroxylases / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D011374:Progesterone; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D013255:Steroid 21-Hydroxylase