title: Olfactory desensitization requires membrane targeting of receptor kinase mediated by beta gamma-subunits of heterotrimeric G proteins.
abstract: Olfaction is mediated by G protein-coupled receptors. In isolated rat olfactory cilia, odorants such as citralva stimulate a burst of cAMP, which peaks in 50 ms and returns almost to base-line level within 150 ms in the continuing presence of odorant. This desensitization is mediated by the cAMP dependent protein kinase and a specialized G protein-coupled receptor kinase originally termed beta ARK2 (GRK3). In vitro experiments suggest that the prenylated beta gamma-subunits of heterotrimeric G proteins target the cytosolic beta ARK1 (GRK2) enzyme to its membrane bound receptor substrate by binding to sites in its carboxyl terminus. Here we demonstrate that odorants stimulate translocation of GRK3 from cytosol to membranes in isolated rat olfactory cilia. We introduced a glutathione S-transferase-GRK3ct fusion protein, containing the carboxyl-terminal 222 amino acid residues of GRK3, which includes the beta gamma binding site, or a 28-amino acid peptide derived therefrom, into permeabilized cilia preparations. These reagents block odorant-mediated enzyme translocation and desensitization while markedly attenuating odorant-stimulated phosphorylation of olfactory proteins. These findings suggest that beta gamma-subunits may physiologically regulate a G protein-coupled receptor kinase and that enzyme translocation may be a general and required feature of the activity of some members of this enzyme family.
authors.affiliation: Department of Zoophysiology, Universität Hohenheim, Stuttgart, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Boekhoff
authors.forename: J
authors.initials: J
authors.lastname: Inglese
authors.forename: S
authors.initials: S
authors.lastname: Schleicher
authors.forename: W J
authors.initials: WJ
authors.lastname: Koch
authors.forename: R J
authors.initials: RJ
authors.lastname: Lefkowitz
authors.forename: H
authors.initials: H
authors.lastname: Breer
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002923:Cilia / Q000502:physiology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D018034:Olfactory Receptor Neurons / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction; D012903:Smell / Q000502:physiology*
chemical_list: D005982:Glutathione Transferase; D011494:Protein Kinases; D019204:GTP-Binding Proteins