Purification and characterization of beta-glucuronidase from bull seminal plasma and its role in fertilization.
Bull seminal plasma contains high levels of beta-glucuronidase. The present study describes the isolation and characterization of beta-glucuronidase, and its role in fertilization. beta-glucuronidase was purified by ion exchange chromatography, saccharolactone-agarose affinity chromatography, and gel filtration. The specific activity of the purified enzyme was 4,414 mumoles/mg protein/min. The purified enzyme showed a single band on 7.5% PAGE. On SDS-PAGE, the enzyme appeared to consist of four identical subunits of M(r) 75,000 each. The apparent Km and Vmax for beta-glucuronidase were 0.4 mM and 5.7 mumol/min using phenolpthalein mono-beta-glucuronic acid as the substrate. beta-glucuronidase appeared to accelerate the cumulus dispersion in vitro.
Rethinaswamy
A
A
Department of Biochemistry, University of Georgia, Athens 30602.
Yang
C H
CH
Srivastava
P N
PN
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D005306:Fertilization / Q000502:physiology*; D005966:Glucuronidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000502:physiology; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D006080:Ovarian Follicle / Q000378:metabolism; D011817:Rabbits; D012661:Semen / Q000201:enzymology*
D005966:Glucuronidase