Hosokawa_1990_Arch.Biochem.Biophys_277_219

Seven carboxylesterase isozymes were purified to electrophoretic homogeneity from liver microsomes of mouse, hamster, guinea pig, rabbit, and monkey by the same procedure used previously to obtain three isozymes from the rat, and their physical, enzymological, and immunological properties were compared with those of the rat isozymes. The substrate specificity and immunological reactivity of liver microsomal carboxylesterases from pig, cow, beagle dog, and human were also examined for comparison, though these enzymes were not purified. The ten purified preparations have similar subunit weight (57,000-64,000), but their isoelectric points differ widely (4.7-6.5). The purification procedure of all isozymes included concanavalin A-Sepharose column chromatography. The isozymes were not eluted from the column with a high concentration of sodium chloride, but were efficiently eluted with alpha-methylmannoside. This observation suggested that the carboxylesterases studied are glycoproteins. All the isozymes except rat RL1 and RL2 possess a high hydrolytic activity toward all the substrates examined. Long-chain monoglyceride was hydrolyzed by the purified carboxylesterase isozymes. Anti-rat RH1 immunoglobulin G was found to possess high cross-reactivity with all isozymes tested, except monkey MK2, by immunoblotting analysis. The amino acid compositions of carboxylesterase isozymes showed considerable similarities, except for monkey MK2. The amino-terminal amino acid sequences showed a striking homology, except for monkey MK2, though the amino-terminal amino acid itself was different in every isozyme. Hepatic microsomal carboxylesterases in mammals play an important role in drug and lipid metabolism in the endoplasmic reticulum, and it is noteworthy that the isozymes from various species examined here showed considerable similarities in physical, enzymatic, and immunochemical properties.