title: Beta-glucuronidase of rat preputial gland.  Crystallization, properties, carbohydrate composition, and subunits.
abstract: Rat preputial gland beta-glucuronidase [ED 3.2.1.31] was purified by ammonium sulfate precipitation, ethanol fractionation, gel filtration on Sephadex G-200 and crystallization.  The purified enzyme appeared homogeneous on electrophoresis in polyacrylamide gel, and on analytical ultracentrifugation and had a molecular weight of approximately 320,000, and a sedimentation coefficient of 12S.  SDS polyacrylamide gel electrophoresis indicated that the enzyme consisted of subunits with molecular weight of 79,000, so the native enzyme appeared to be a tetramer.  The Km with p-nitrophenyl beta-D-glucosiduronic acid as substrate was about 0.53 mM.  The enzyme had a single pH optimum at 4.5.  The enzyme had a very low content of sulphur-containing amino acid and contained 5.7 per cent carbohydrate, consisting of mannose, glucose, fucose, galactose, and glucosamine in a ratio of 44;9;6;2;41.  Sialic acid was not detected in the crystallized enzyme.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002987:Clitoris / Q000201:enzymology*; D003460:Crystallization; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005966:Glucuronidase / Q000302:isolation & purification / Q000378:metabolism; D006595:Hexosamines / Q000032:analysis; D006601:Hexoses / Q000032:analysis; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D051381:Rats; D012627:Sebaceous Glands / Q000201:enzymology*; D012710:Serum Albumin, Bovine; D012794:Sialic Acids / Q000032:analysis
chemical_list: D000596:Amino Acids; D006595:Hexosamines; D006601:Hexoses; D046911:Macromolecular Substances; D012794:Sialic Acids; D012710:Serum Albumin, Bovine; D005966:Glucuronidase