title: The gamma-aminobutyric acid receptor B, but not the metabotropic glutamate receptor type-1, associates with lipid rafts in the rat cerebellum.
abstract: Recent evidence suggests that specialized microdomains, called lipid rafts, exist within plasma membranes. These domains are enriched in cholesterol and sphingolipids and are resistant to non-ionic detergent-extraction at 4 degrees C. They contain specific populations of membrane proteins, and can change their size and composition in response to cellular signals, resulting in activation of signalling cascades. Here, we demonstrate that both the metabotropic gamma-aminobutyric acid receptor B (GABA(B) receptor) and the metabotropic glutamate receptor-1 from rat cerebellum are insoluble in the non-ionic detergent Triton X-100. However, only the GABA(B) receptor associates with raft fractions isolated from rat brain by sucrose gradient centrifugation. Moreover, increasing the stringency of isolation by decreasing the protein : detergent ratio caused an enrichment of the GABA(B) receptor in raft fractions. In contrast, depletion of cholesterol from cerebellar membranes by either saponin or methyl-beta-cyclodextrin treatment, which solubilize known raft markers, also increased the solubility of the GABA(B) receptor. These properties are all consistent with an association of the GABA(B) receptor with lipid raft microdomains.
authors.affiliation: Medical Research Council Anatomical Neuropharmacology Unit, Mansfield Road, Oxford, UK.
authors.forename: A
authors.initials: A
authors.lastname: Becher
authors.forename: J H
authors.initials: JH
authors.lastname: White
authors.forename: R A
authors.initials: RA
authors.lastname: McIlhinney
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000737:chemistry; D002499:Centrifugation, Density Gradient; D002531:Cerebellum / Q000737:chemistry* / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry; D003505:Cyclodextrins / Q000737:chemistry / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000737:chemistry; D006017:Glycolipids / Q000737:chemistry; D021962:Membrane Microdomains / Q000737:chemistry* / Q000378:metabolism; D017830:Octoxynol / Q000737:chemistry / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000187:drug effects; D021122:Protein Subunits; D051381:Rats; D017208:Rats, Wistar; D018080:Receptors, GABA-B / Q000737:chemistry* / Q000378:metabolism; D018094:Receptors, Metabotropic Glutamate / Q000737:chemistry*; D012503:Saponins / Q000737:chemistry / Q000494:pharmacology; D012995:Solubility / Q000187:drug effects; D047392:beta-Cyclodextrins
chemical_list: D003505:Cyclodextrins; D006017:Glycolipids; D021122:Protein Subunits; D018080:Receptors, GABA-B; D018094:Receptors, Metabotropic Glutamate; D012503:Saponins; D047392:beta-Cyclodextrins; C104077:metabotropic glutamate receptor type 1; D017830:Octoxynol; D002784:Cholesterol; D019204:GTP-Binding Proteins; C031215:betadex