Multisite contacts involved in coupling of the beta-adrenergic receptor with the stimulatory guanine-nucleotide-binding regulatory protein. Structural and functional studies by beta-receptor-site-specific synthetic peptides.
Synthetic peptides, 12-22 amino acid residues long, comprising the presumed coupling sites of the beta-adrenergic receptor with the stimulatory guanine-nucleotide-binding regulatory protein (Gs), were examined for their ability to modulate Gs activation in turkey erythrocyte membranes. Three peptides corresponding to the second cytoplasmic loop, the N-terminal region of the third cytoplasmic loop, and the N-terminal region of the putative fourth cytoplasmic loop, compete synergistically with the hormone-stimulated receptor for Gs activation with median effector concentrations of 15-35 microM, or 3-4 microM for combinations of two peptides. One peptide, corresponding to the C-terminal region of the third cytoplasmic loop, carries the unique ability to activate the Gs-adenylate-cyclase complex independent of the signalling state of the receptor. These observations are consistent with a dynamic model of receptor-mediated G-protein activation in membranes, where domains composed of the second, third and fourth intracellular loop of the receptor bind to and are interactive with the G-protein heterotrimer, resulting in ligand-induced conformational changes of the receptor. In response to hormone binding, the extent or the number of sites involved in interaction with Gs may be readjusted using a fourth site. Modulation of coupling sites may elicit congruent conformational changes within the Gs heterotrimer, with qualitatively different effects on GTP/GDP exchange in the alpha subunit of Gs and downstream effector regulation. This model corroborates and expands a similar model suggested for activated rhodopsin-transducin interaction [König, B., Arendt, A., McDowell, J. H., Kahlert, M., Hargrave, P. A. & Hofmann, K. P. (1989) Proc. Natl Acad. Sci. USA 86, 6878-6882].
Münch
G
G
Physiologisch-Chemisches Institut, Universität Würzburg, Federal Republic of Germany.
Dees
C
C
Hekman
M
M
Palm
D
D
D000535:Aluminum / Q000494:pharmacology; D017607:Aluminum Compounds; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D004082:Dihydroalprenolol / Q000097:blood; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005459:Fluorides; D005461:Fluorine / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000097:blood*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D011487:Protein Conformation; D011943:Receptors, Adrenergic, beta / Q000187:drug effects / Q000378:metabolism*; D014422:Turkeys
D017607:Aluminum Compounds; D009842:Oligopeptides; D010455:Peptides; D011943:Receptors, Adrenergic, beta; C050992:tetrafluoroaluminate; D005461:Fluorine; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D004082:Dihydroalprenolol; D000535:Aluminum; D019204:GTP-Binding Proteins; D005459:Fluorides