Multiplicity and regulation of hepatic microsomal carboxylesterases in rats.
Three isozymes of carboxylesterase were purified from rat liver microsomes by using Sephadex G-150 gel filtration, DE-52 ion exchange, and chromatofocusing column chromatographies. These isozymes each showed a single protein band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were immunologically different from each other as determined by immunochemical blotting analysis and immunochemical inhibition of catalytic activity. The three isozymes were named RH1 (molecular weight 174,000, trimer, pl 6.0), RL1 (molecular weight 61,000, monomer, pl 6.5), and RL2 (molecular weight 61,000, monomer, pl 5.5). RL1 has the highest specific activities toward p-nitrophenylacetate and malathion. Acetanilide is a rather specific substrate for RL2, whereas RH1 has the highest specific activity for butanilicaine. RL1 has the highest specific activity for the hydrolysis of long-chain acyl-CoA. To investigate the hormonal regulation of carboxylesterase activities, we have quantitated RL1, RL2, and RH1 in liver microsomes from male and female rats using a radial immunodiffusion assay. The amount of RL1 in male rats was decreased by castration but recovered to almost the level in sham-operated rat liver microsomes after treatment of the castrated rats with testosterone. Conversely, in ovariectomized female rats, the amount of RL1 was increased as compared to that in sham-operated rats, and treatment of the ovariectomized rats with estradiol tended to decrease the quantity of RL1. In all cases of sex hormone treatment, the amount of RH1 remains unclear at present. However, the amount of RL2 may be, at least in part, regulated by estrogens. On the other hand, phenobarbital treatment of male and female rats caused a significant increase in the amounts of RH1 and RL2, whereas RL1 was not affected. It was concluded that the three isozymes differ considerably from each other in response to hormone treatment, inducibility, substrate specificity, and immunological properties.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats; D011919:Rats, Inbred Strains; D012737:Sex Factors; D013379:Substrate Specificity
D000906:Antibodies; D000936:Antigen-Antibody Complex; D007527:Isoenzymes; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase