title: Activation of cardiac G-proteins by muscarinic acetylcholine receptors: regulation by Mg2+ and Na+ ions.
abstract: The mechanism of G-protein activation mediated by cardiac muscarinic acetylcholine receptors (mAcChR) was studied in porcine atrial membranes by measuring the agonist-stimulated hydrolysis of GTP and G-protein GTPase activity. The cholinergic agonist, carbachol, stimulated a high-affinity GTPase activity in mAcChR-enriched membranes by maximally about 70%. Carbachol increased the Vmax of GTP hydrolysis without affecting the Km for GTP (approximately 0.2 microM). The activity of low-affinity GTPase(s) was not altered by carbachol. Carbachol and the mAcChR agonist oxotremorine induced the same maximal response, with half-maximal activation occurring at about 5 and 0.5 microM, respectively. The stimulatory effect of carbachol was blocked by the mAcChR antagonist, atropine. The stimulation of GTP hydrolysis by cholinergic agonist, was strictly dependent on Mg2+, with half-maximal stimulation observed at 4-5 microM free Mg2+. The monovalent cations, in the potency order, Na+ greater than Li+ greater than K+, enhanced the GTP hydrolysis stimulated by carbachol at a maximally effective concentration. The increase in the maximal agonist response seen with NaCl was the result of NaCl decreasing the basal more than the carbachol-stimulated GTP hydrolysis. The data indicate that occupation of mAcChR in porcine atrial membranes by agonists leads to an increase in G-protein GTPase activity, apparently by increasing the amount of nucleotide-free G-protein to which GTP can bind without altering the apparent binding affinity for GTP. This receptor-induced G-protein activation is under the strict control of Mg2+ acting at low, microM concentrations, and is additionally modulated by monovalent cations.
authors.affiliation: Pharmakologisches Institut, Universität Heidelberg, F.R.G.
authors.forename: G
authors.initials: G
authors.lastname: Hilf
authors.forename: K H
authors.initials: KH
authors.lastname: Jakobs
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology*; D008566:Membranes / Q000187:drug effects / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology; D011813:Quinuclidinyl Benzilate; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000378:metabolism*; D012964:Sodium / Q000494:pharmacology*; D012965:Sodium Chloride / Q000494:pharmacology; D013552:Swine; D010566:Virulence Factors, Bordetella / Q000378:metabolism
chemical_list: D011976:Receptors, Muscarinic; D010566:Virulence Factors, Bordetella; D012965:Sodium Chloride; D000244:Adenosine Diphosphate; D011813:Quinuclidinyl Benzilate; D002217:Carbachol; D012964:Sodium; D000251:Adenosine Triphosphatases; D019204:GTP-Binding Proteins; D008274:Magnesium