title: Specific enhancement of beta-adrenergic receptor kinase activity by defined G-protein beta and gamma subunits.
abstract: The beta and gamma subunits of heterotrimeric guanine nucleotide-binding proteins (G proteins) have recently been shown to play an active role in signal transduction. Among other effects they enable translocation of the beta-adrenergic receptor kinase (beta ARK) from the cytosol to the plasma membrane and thus permit phosphorylation and ultimately desensitization of beta-adrenergic receptors and other G-protein-coupled receptors. To investigate the specificity of this effect, we have purified various combinations of recombinant beta and gamma subunits expressed in Sf9 cells and measured their effects on beta ARK-catalyzed phosphorylation of beta 2-adrenergic receptors and of rhodopsin. The combinations tested were beta 1 gamma 2, beta 1 gamma 3, beta 2 gamma 2, beta 2 gamma 3, and transducin beta gamma (beta 1 gamma 1). There were clear differences in enhancement of rhodopsin phosphorylation, with an order of efficacy beta 2 gamma 2 > beta 1 gamma 2 >> beta 2 gamma 3 approximately beta 1 gamma 3 approximately beta 1 gamma 1. The first two combinations had larger effects than a mixed beta gamma preparation from bovine brain. In enhancing phosphorylation of beta 2-adrenergic receptors, beta 1 gamma 2 was more efficient and potent than all other combinations. These data suggest a twofold specificity of beta gamma complexes in enhancing beta ARK-catalyzed receptor phosphorylation: the gamma subunits may be important in interacting with beta ARK, with gamma 2 being more potent than gamma 3, whereas the beta subunits may determine coupling to the receptors, with beta 2 being more effective than beta 1 for rhodopsin and beta 1 being more effective than beta 2 for beta 2-adrenergic receptors.
authors.affiliation: Laboratory of Molecular Biology, University of Munich, Max Planck Institute of Biochemistry, Martinsried, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Müller
authors.forename: M
authors.initials: M
authors.lastname: Hekman
authors.forename: M J
authors.initials: MJ
authors.lastname: Lohse
mesh_terms: D000818:Animals; D016367:Baculoviridae; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D009036:Moths; D010750:Phosphoproteins / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012243:Rhodopsin / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D051552:beta-Adrenergic Receptor Kinases
chemical_list: D010750:Phosphoproteins; D011943:Receptors, Adrenergic, beta; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D012243:Rhodopsin; D017868:Cyclic AMP-Dependent Protein Kinases; D051552:beta-Adrenergic Receptor Kinases; D019204:GTP-Binding Proteins