Purification and properties of a form of UDP-glucuronyltransferase from liver microsomes of 3-methylcholanthrene-treated rats.
A form of UDP-glucuronyltransferase has been purified from liver microsomes of 3-methylcholanthrene-treated rats by a simple and rapid method involving chromatography on DEAE-Toyopearl and UDP-hexanolamine Sepharose columns. The purified preparation gave a single protein band (Mr 54,000) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It catalyzed the glucuronidation of not only phenolic xenobiotics such as 4-nitrophenol, 1-naphthol, and eugenol but also serotonin, which is an endogenous compound. Its activities toward 4-hydroxybiphenyl and testosterone were very low and no activity was detected toward bilirubin. After removal of the detergent (Emulgen 911), the transferase activity was stimulated by various phospholipids, about 10-fold activation being attained with phosphatidylcholine and lysophosphatidylcholine. On nitrocellulose sheets concanavalin A, but not wheat germ agglutinin, bound to the purified transferase, and this binding was abolished in the presence of alpha-methylmannoside and after treatment of the enzyme with endo-beta-N-acetylglucosaminidase H (Endo H). These observations provided evidence that the transferase is a glycoprotein carrying a "high mannose type" of oligosaccharide chain(s). The NH2-terminal 7 residues of the purified enzyme were determined to be Thr-Lys-Leu-Leu-Val-Trp-Pro.
Yokota
H
H
Department of Veterinary Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Hokkaido.
Yuasa
A
A
Sato
R
R
D000818:Animals; D002845:Chromatography / Q000379:methods; D014453:Glucuronosyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D037102:Lectins; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology*; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity
D037102:Lectins; D008748:Methylcholanthrene; D014453:Glucuronosyltransferase