Purification and characterization of a cytochrome P-450 isozyme isolated from the liver of rats pretreated with clofibrate.
1- Using a methodology of purification consisting of only one chromatographic step (phenyl-Sepharose) we have purified a cytochrome P-450 isozyme from liver of clofibrate - treated rats. It was called cytochrome P-450 clo. 2- A single polypeptide of mol.wt. 50,000 was visible after sodium dodecyl sulphate polyacrylamide gel electrophoresis. 3- Antiserum raised against the pure enzyme specifically recognized P-450 clo and inhibited more than 90% of the 11- and 12- laurate hydroxylase activities present in clofibrate - treated rats. 4- Clofibrate treatment of the rats resulted in a six fold increase in microsomal cytochrome P-450 clo as judged by immunochemical quantification. This result is in agreement with the increase of laurate hydroxylase activity after treatment by clofibrate.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex / Q000032:analysis; D002994:Clofibrate / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D005779:Immunodiffusion; D007527:Isoenzymes / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D000906:Antibodies; D000936:Antigen-Antibody Complex; D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System; D002994:Clofibrate