title: Purification to apparent homogeneity and partial characterization of rat liver UDP-glucose:glycoprotein glucosyltransferase.
abstract: The UDP-Glc:glycoprotein glucosyltransferase is a soluble protein of the endoplasmic reticulum that catalyzes the glucosylation of protein-linked, glucose-free, high mannose-type oligosaccharides. In vivo, the newly glucosylated compounds are immediately deglucosylated, presumably by glucosidase II. The glucosyltransferase has been purified to apparent homogeneity from rat liver. The enzyme appears to have a molecular weight of 150,000 and 270,000 under denaturing and native conditions, respectively. The pure enzyme shows an almost absolute requirement for Ca2+ ions and for UDP-Glc as sugar donor. The same as crude preparations, the pure enzyme synthesized Glc1 Man7-9GlcNAc2-protein from Man7-9GlcNAc2-protein. Denatured glycoproteins are glucosylated much more efficiently than native ones by the apparently homogeneous glucosyltransferase. Availability of the pure enzyme will allow testing the possible involvement of transient glucosylation of glycoproteins in the folding of glycoproteins and/or in the mechanism by which cells dispose of malfolded glycoproteins in the endoplasmic reticulum.
authors.affiliation: Instituto de Investigaciones Bioquímicas Fundación Campomar, Buenos Aires, Argentina.
authors.forename: S E
authors.initials: SE
authors.lastname: Trombetta
authors.forename: A J
authors.initials: AJ
authors.lastname: Parodi
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005964:Glucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D051381:Rats
chemical_list: D005964:Glucosyltransferases; C072423:mannosylglycoprotein 1,3-glucosyltransferase