title: Cryptococcus laurentii cell envelope glycoprotein. Evidence for separate oligosaccharide side chains of different composition and structure.
abstract: Particulate enzyme preparations of the fungus imperfectus Cryptococcus laurentii catalyze transfer of mannosyl and galactosyl residues from GDP-[14C]mannose and UDP-[3H]-galactose to the same endogenous acceptor. After solubilization with pronase, the major portion of both labels is retarded on Sepharose columns and forms a symmetrical peak, in which 14C and 3H coincide. Label also coincides with endogenous protein and carbohydrate. Both labels bind to Sepharose-Concanavalin A (Con A) and are eluted with alpha-methylglucoside. After beta elimination with NaOH-NaBH4 only 14C label retains binding to Sepharose-Con A; 3H label representing (6-O-alpha-galactosyl)10-O-beta-galactosyl-O-mannitol as previously reported (Raizada, M. K., Kloepfer, H. G., Schutzbach, J. S., and Ankel, H. (1974) J. Biol. Chem. 249, 6080-6086) no longer binds. The [14C]mannose-containing material after beta elimination yields a pentasaccharide and a trisaccharide. Similar penta- and trisaccharides can be isolated following beta elimination of particulate preparations of the organism after pronase treatment. Analytical data suggest that the structure of the isolated pentasaccharides corresponds to that of a pentasaccharide previously synthesized de novo using cell-free enzyme preparations of the organism: 2-O-alpha-mannosyl-6-O-alpha-mannosyl-3-O-alpha-mannosyl-(2-O-beta-xylosyl)-O-mannose (Schutzbach, J. S., Raizada, M. K., and Ankel, H. (1974) J. Biol. Chem. 249, 2953-2958). The trisaccharide has the structure 2-O-alpha-mannosyl-2-O-alpha-mannosyl-O-mannitol. The data are consistent with a glycoprotein structure in which these three types of oligosaccharides are bound to a common polypeptide core through O-glycosidic linkages to threonyl and seryl residues.
mesh_terms: D002462:Cell Membrane / Q000032:analysis*; D002847:Chromatography, Agarose; D003208:Concanavalin A; D003454:Cryptococcus / Q000032:analysis*; D006023:Glycoproteins / Q000032:analysis*; D006155:Guanosine Diphosphate Mannose / Q000378:metabolism; D008968:Molecular Conformation; D009844:Oligosaccharides / Q000302:isolation & purification*; D010455:Peptides; D014531:Uridine Diphosphate Galactose / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D009844:Oligosaccharides; D010455:Peptides; D003208:Concanavalin A; D014531:Uridine Diphosphate Galactose; D006155:Guanosine Diphosphate Mannose