title: The preparation of rat liver lysosome membranes. Several membrane proteins contain complex oligosaccharides.
abstract: Lysosome membranes were isolated, and membrane proteins and glycoproteins were characterized by electrophoresis and lectin probes of nitrocellulose blots. Rat liver lysosomes were isolated on a discontinuous metrizamide gradient and characterized by subcellular marker enzymes. Lysosomes were lysed by hypotonic freeze-thaw shock and membranes were isolated. The release of beta-N-acetylhexosaminidase was used to monitor the disruption of the lysosomes. Proteins of lysosome membranes were analyzed by sodium dodecyl sulfate - polyacrylamide gel electrophoresis. There were at least 30 proteins present and several were glycoproteins. Nitrocellulose blots of lysosome membrane proteins were probed with a panel of lectins, including concanavalin A, Ulex europaeus agglutinin I, Lotus tetragonolobus agglutinin, soybean agglutinin, peanut agglutinin, and Ricinus communis agglutinin I. Peanut agglutinin and Ricinus communis agglutinin I binding were also examined after neuramidase treatment of lysosome membranes. Ten proteins bound concanavalin A, and neuraminidase pretreatment revealed six proteins that bound Ricinus communis agglutinin I and three proteins that bound peanut agglutinin. The other lectins tested did not bind to any lysosome membrane proteins. These results indicate that lysosome membranes contain several glycoproteins, some of which contain sialic acid terminating complex oligosaccharides.
authors.affiliation: Department of Biochemistry, Children's Psychiatric Research Institute, London, Ont., Canada.
authors.forename: C A
authors.initials: CA
authors.lastname: Rupar
authors.forename: J D
authors.initials: JD
authors.lastname: Whitehall
mesh_terms: D000818:Animals; D007425:Intracellular Membranes / Q000032:analysis*; D008099:Liver / Q000032:analysis*; D008247:Lysosomes / Q000032:analysis* / Q000201:enzymology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis*; D009844:Oligosaccharides / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D008565:Membrane Proteins; D009844:Oligosaccharides