title: Domains of yeast plasma membrane and ATPase-associated glycoprotein.
abstract: In yeast homogenates the plasma membrane H(+)-ATPase and a major surface glycoprotein of about 115 kDa are present in two membrane fractions with peak densities in sucrose gradients of 1.17 and 1.22. Immunogold electron microscopy of frozen yeast sections indicates that the ATPase is exclusively (greater than 95%) present at the surface membrane. Therefore the two ATPase-containing fractions appear to correspond to different domains of the plasma membrane. The 115 kDa glycoprotein is tightly associated with the ATPase during solubilization and purification of the enzyme. However, in a mutant lacking the glycoprotein the activity of the plasma membrane H(+)-ATPase is similar to wild type, suggesting that this association is fortuitous. The ATPase and the glycoprotein are difficult to separate by electrophoresis and therefore binding of concanavalin A to the ATPase cannot be unambiguously demonstrated in wild-type yeast. By utilizing the mutant without glycoprotein it was shown that the ATPase band of 105 kDa binds concanavalin A.
authors.affiliation: European Molecular Biology Laboratory, Heidelberg, F.R.G.
authors.forename: R
authors.initials: R
authors.lastname: Serrano
authors.forename: C
authors.initials: C
authors.lastname: Montesinos
authors.forename: M
authors.initials: M
authors.lastname: Roldán
authors.forename: G
authors.initials: G
authors.lastname: Garrido
authors.forename: C
authors.initials: C
authors.lastname: Ferguson
authors.forename: K
authors.initials: K
authors.lastname: Leonard
authors.forename: B C
authors.initials: BC
authors.lastname: Monk
authors.forename: D S
authors.initials: DS
authors.lastname: Perlin
authors.forename: E W
authors.initials: EW
authors.lastname: Weiler
mesh_terms: D002462:Cell Membrane / Q000737:chemistry / Q000201:enzymology; D002500:Centrifugation, Isopycnic; D005656:Fungal Proteins / Q000032:analysis* / Q000235:genetics; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics; D016253:Microscopy, Immunoelectron; D009154:Mutation; D006180:Proton-Translocating ATPases / Q000032:analysis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000235:genetics / Q000648:ultrastructure
chemical_list: D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D006180:Proton-Translocating ATPases