title: Enzymatic properties of human glucuronyltransferase and a sensitive method for its assay in a stable B lymphocyte cell line.
abstract: Properties of lymphocyte glucuronyltransferase were studied in homogenates of SN1006 cells. A sensitive assay procedure for lymphocyte glucuronyltransferase was developed utilizing radioactive testosterone as the acceptor substrate and TLC for separation of the metabolite. The method is capable of detecting picomolar quantities of the product. The enzyme activity exhibited a broad pH optimum, and was subject to activation by the detergent Lubrol WX and Mn++ ions. The activity conformed to the Michaelis-Menten kinetics giving apparent Km values of 0.8 mM and 11 microM, for UDPGA and testosterone, respectively. 4-Methylumbelliferone, a-naphthol and p-nitrophenol behaved as competitive inhibitors of testosterone glucuronidation. The results indicate that the method could be used for genetic studies of human lymphocyte glucuronyltransferase, and that the enzyme is of consequence in detoxication of exogenous as well as endogenous substrates.
mesh_terms: D001402:B-Lymphocytes / Q000201:enzymology; D002460:Cell Line; D003470:Culture Media; D004789:Enzyme Activation; D014453:Glucuronosyltransferase / Q000037:antagonists & inhibitors / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008345:Manganese / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D013739:Testosterone / Q000378:metabolism
chemical_list: D003470:Culture Media; C017474:Lubrol WX; D011092:Polyethylene Glycols; D013739:Testosterone; D008345:Manganese; D014453:Glucuronosyltransferase