title: Organization of G proteins and adenylyl cyclase at the plasma membrane.
abstract: There is mounting evidence for the organization and compartmentation of signaling molecules at the plasma membrane. We find that hormone-sensitive adenylyl cyclase activity is enriched in a subset of regulatory G protein-containing fractions of the plasma membrane. These subfractions resemble, in low buoyant density, structures of the plasma membrane termed caveolae. Immunofluorescence experiments revealed a punctate pattern of G protein alpha and beta subunits, consistent with concentration of these proteins at distinct sites on the plasma membrane. Partial coincidence of localization of G protein alpha subunits with caveolin (a marker for caveolae) was observed by double immunofluorescence. Results of immunogold electron microscopy suggest that some G protein is associated with invaginated caveolae, but most of the protein resides in irregular structures of the plasma membrane that could not be identified morphologically. Because regulated adenylyl cyclase activity is present in low-density subfractions of plasma membrane from a cell type (S49 lymphoma) that does not express caveolin, this protein is not required for organization of the adenylyl cyclase system. The data suggest that hormone-sensitive adenylyl cyclase systems are localized in a specialized subdomain of the plasma membrane that may optimize the efficiency and fidelity of signal transduction.
authors.affiliation: Department of Pharmacology, The University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75235-9041, USA.
authors.forename: C
authors.initials: C
authors.lastname: Huang
authors.forename: J R
authors.initials: JR
authors.lastname: Hepler
authors.forename: L T
authors.initials: LT
authors.lastname: Chen
authors.forename: A G
authors.initials: AG
authors.lastname: Gilman
authors.forename: R G
authors.initials: RG
authors.lastname: Anderson
authors.forename: S M
authors.initials: SM
authors.lastname: Mumby
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D000918:Antibody Specificity; D051242:Caveolin 1; D022461:Caveolins; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D004285:Dogs; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D020962:Heterotrimeric GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D016253:Microscopy, Immunoelectron; D011485:Protein Binding; D013347:Subcellular Fractions / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure
chemical_list: C495540:CAV1 protein, human; C495541:Cav1 protein, mouse; D051242:Caveolin 1; D022461:Caveolins; D008565:Membrane Proteins; D020962:Heterotrimeric GTP-Binding Proteins; D000262:Adenylyl Cyclases