Studies on the glycosidases of semen. Further purification and characterization of two hexosaminidases from bull seminal plasma.
Two isozymes of beta-N-acetylglucosaminidase (2-acetamido-2-deoxy-beta-D-glucoside acetamidodeoxy glucohydrolase, EC 3.2.1.30) (A and B) from bull seminal plasma were purified to homogeneity by isoelectric focusing having pI values of 5.31 and 6.78. The two proteins were glycoproteins with very similar amino acid composition but isozyme A contained more sialic acid than isozyme B. The molecular weights of isozyme A and B were estimated at 200 000 and 190 000 by gel filtration. Two identical subunits corresponding to molecular weights of 53 000 and 13 400 were obtained from hexosaminidase A and B when subjected to polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. Similar results were obtained when dissociation of the isozymes was effected with mercaptoethanol, guanidine hydrochloride and urea in presence of sodium dodecyl sulphate and the subunits separated by acrylamide gel electrophoresis. The two isozymes were more stable in frozen conditions than at the refrigerated temperature. Of the divalent ion tested, glucosaminidase and galactosaminidase activities of isozymes A and B were strongly inhibited by Hg2+ and Ag+ thus suggesting the presence of thiol groups in the two proteins. The two isozymes were active on natural substrates; isozyme B being more active than isozyme A.
D000118:Acetylglucosaminidase / Q000302:isolation & purification / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002686:Chitin / Q000378:metabolism; D006020:Glycopeptides / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008297:Male; D008628:Mercury / Q000494:pharmacology; D008970:Molecular Weight; D012661:Semen / Q000201:enzymology*; D012834:Silver / Q000494:pharmacology; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D000596:Amino Acids; D006020:Glycopeptides; D007527:Isoenzymes; D013439:Sulfhydryl Reagents; D002686:Chitin; D012834:Silver; D006596:Hexosaminidases; D000118:Acetylglucosaminidase; D008628:Mercury