title: Evidence against a regulation of Na+/K(+)-ATPase by Gi proteins. Failure to detect an influence of G proteins on Na+/Ca(2+)-exchange in cardiac sarcolemmal membranes.
abstract: In the myocardium the inhibitory guanine nucleotide-binding regulatory proteins (Gi proteins) mediate negative chronotropic and negative inotropic effects by activation of K+ channels and inhibition of adenylyl cyclase. The concept of a uniform inhibitory action of Gi proteins on myocardial cellular activity has been questioned by the recent observations of adenosine-induced activation of the Na+/Ca(2+) exchange and a carbachol-induced inhibition of the Na+/K(+)-ATPase activity in cardiac sarcolemmal membranes. The aim of the present study, therefore, was to reinvestigate the putative regulation of Na+/Ca(2+) exchange and Na+/K(+)-ATPase activity in purified canine sarcolemmal membranes. These membranes were enriched in adenosine A1 (Maximum number of receptors, Bmax 0.033 pmol/mg) and muscarinic M2 (Bmax 2.9 pmol/mg) receptors and contained Gi2 and Gi3, two Gi protein isoforms, and G0, another pertussis toxin-sensitive G protein, as detected with specific antibodies. The adenosine A1-selective agonist, (-)-N6-(2-phenylisopropyl)-adenosine, and the muscarinic agonist, carbachol, both inhibited isoprenaline-stimulated adenylyl cyclase activity by 25% and 35% respectively, and the stable GTP analogue 5'-guanylylimidodiphosphate inhibited forskolin-stimulated adenylyl cyclase activity by 35% in these membranes. The characteristics of Na+/Ca(2+) exchange and Na+/K(+)-ATPase activity as well as those of the ouabain-sensitive, K(+)-activated 4-nitrophenylphosphatase, an ATP-independent, partial reaction of the Na+/K(+)-ATPase, were in agreement with published data with regard to specific activity, time course of activity and substrate dependency. However, none of these activities were influenced by adenosine, (-)-N6-(2-phenylisopropyl)-adenosine, carbachol, or stable GTP analogs, suggesting that Na+/Ca(2+) exchange and Na+/K(+)-ATPase are not regulated by Gi proteins in canine cardiac sarcolemmal membranes.
authors: Universität Heidelberg, Innere Medizin III, Germany.
authors: R A
authors: RA
authors: Mura
authors: F
authors: F
authors: Zeifang
authors: L
authors: L
authors: Piacentini
authors: W
authors: W
authors: Kübler
authors: B
authors: B
authors: Rauch
authors: F
authors: F
authors: Niroomand
mesh_terms: D009597:4-Nitrophenylphosphatase / Q000378:metabolism; D000067956:Adenylyl Cyclase Inhibitors; D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D002352:Carrier Proteins / Q000187:drug effects* / Q000378:metabolism; D004285:Dogs; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D018721:Muscarinic Agonists / Q000494:pharmacology; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000166:cytology / Q000201:enzymology; D010660:Phenylisopropyladenosine / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000378:metabolism; D018047:Receptors, Purinergic P1 / Q000378:metabolism; D012508:Sarcolemma / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D019831:Sodium-Calcium Exchanger; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism*
chemical_list: D000067956:Adenylyl Cyclase Inhibitors; D002352:Carrier Proteins; D018721:Muscarinic Agonists; D015221:Potassium Channels; D011976:Receptors, Muscarinic; D018047:Receptors, Purinergic P1; D019831:Sodium-Calcium Exchanger; D010660:Phenylisopropyladenosine; D006165:Guanylyl Imidodiphosphate; D006160:Guanosine Triphosphate; D002217:Carbachol; D012964:Sodium; D009597:4-Nitrophenylphosphatase; D019204:GTP-Binding Proteins; D000254:Sodium-Potassium-Exchanging ATPase; D002118:Calcium