Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1993 Mar;91(3):939–949. doi: 10.1172/JCI116315

Downregulation of cardiac guanosine 5'-triphosphate-binding proteins in right atrium and left ventricle in pacing-induced congestive heart failure.

D A Roth 1, K Urasawa 1, G A Helmer 1, H K Hammond 1
PMCID: PMC288046  PMID: 8383705

Abstract

The extent to which congestive heart failure (CHF) is dependent upon increased levels of the cardiac inhibitory GTP-binding protein (Gi), and the impact of CHF on the cardiac stimulatory GTP-binding protein (Gs) and mechanisms by which Gs may change remain unexplored. We have addressed these unsettled issues using pacing-induced CHF in pigs to examine physiological, biochemical, and molecular features of the right atrium (RA) and left ventricle (LV). CHF was associated with an 85 +/- 20% decrease in LV segment shortening (P < 0.001) and a 3.5-fold increase (P = 0.006) in the ED50 for isoproterenol-stimulated heart rate responsiveness. Myocardial beta-adrenergic receptor number was decreased 54% in RA (P = 0.004) and 57% in LV (P < 0.001), and multiple measures of adenylyl cyclase activity were depressed 49 +/- 8% in RA (P < 0.005), and 44 +/- 9% in LV (P < 0.001). Quantitative immunoblotting established that Gi and Gs were decreased in RA (Gi: 59% reduction; P < 0.0001; Gs: 28% reduction; P < 0.007) and LV (Gi: 35% reduction; P < 0.008; Gs: 28% reduction; P < 0.01) after onset of CHF. Reduced levels of Gi and Gs were confirmed by ADP ribosylation studies, and diminished function of Gs was established in reconstitution studies. Steady state levels for Gs alpha mRNA were increased in RA and unchanged in LV, and significantly more GS alpha was found in the supernatant (presumably cytosolic) fraction in RA and LV membrane homogenates after CHF, suggesting that increased Gs degradation, rather than decreased Gs synthesis, is the mechanism by which Gs is downregulated. We conclude that cardiac Gi content poorly predicts adrenergic responsiveness or contractile function, that decreased Gs is caused by increased degradation rather than decreased synthesis, and that alterations in beta-adrenergic receptors, adenylyl cyclase, and GTP-binding proteins are uniform in RA and LV in this model of congestive heart failure.

Full text

PDF
939

Images in this article

944Image
on p.944
944Image
on p.944
944Image
on p.944
945Image
on p.945
945Image
on p.945
945Image
on p.945
947Image
on p.947

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alousi A. A., Jasper J. R., Insel P. A., Motulsky H. J. Stoichiometry of receptor-Gs-adenylate cyclase interactions. FASEB J. 1991 Jun;5(9):2300–2303. doi: 10.1096/fasebj.5.9.1650314. [DOI] [PubMed] [Google Scholar]
  2. Armstrong P. W., Stopps T. P., Ford S. E., de Bold A. J. Rapid ventricular pacing in the dog: pathophysiologic studies of heart failure. Circulation. 1986 Nov;74(5):1075–1084. doi: 10.1161/01.cir.74.5.1075. [DOI] [PubMed] [Google Scholar]
  3. Bers D. M. Isolation and characterization of cardiac sarcolemma. Biochim Biophys Acta. 1979 Jul 19;555(1):131–146. doi: 10.1016/0005-2736(79)90078-6. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brann M. R., Collins R. M., Spiegel A. Localization of mRNAs encoding the alpha-subunits of signal-transducing G-proteins within rat brain and among peripheral tissues. FEBS Lett. 1987 Sep 28;222(1):191–198. doi: 10.1016/0014-5793(87)80218-1. [DOI] [PubMed] [Google Scholar]
  6. Bristow M. R., Ginsburg R., Umans V., Fowler M., Minobe W., Rasmussen R., Zera P., Menlove R., Shah P., Jamieson S. Beta 1- and beta 2-adrenergic-receptor subpopulations in nonfailing and failing human ventricular myocardium: coupling of both receptor subtypes to muscle contraction and selective beta 1-receptor down-regulation in heart failure. Circ Res. 1986 Sep;59(3):297–309. doi: 10.1161/01.res.59.3.297. [DOI] [PubMed] [Google Scholar]
  7. Böhm M., Gierschik P., Jakobs K. H., Pieske B., Schnabel P., Ungerer M., Erdmann E. Increase of Gi alpha in human hearts with dilated but not ischemic cardiomyopathy. Circulation. 1990 Oct;82(4):1249–1265. doi: 10.1161/01.cir.82.4.1249. [DOI] [PubMed] [Google Scholar]
  8. Calderone A., Bouvier M., Li K., Juneau C., de Champlain J., Rouleau J. L. Dysfunction of the beta- and alpha-adrenergic systems in a model of congestive heart failure. The pacing-overdrive dog. Circ Res. 1991 Aug;69(2):332–343. doi: 10.1161/01.res.69.2.332. [DOI] [PubMed] [Google Scholar]
  9. Coleman H. N., 3rd, Taylor R. R., Pool P. E., Whipple G. H., Covell J. W., Ross J., Jr, Braunwald E. Congestive heart failure following chronic tachycardia. Am Heart J. 1971 Jun;81(6):790–798. doi: 10.1016/0002-8703(71)90083-4. [DOI] [PubMed] [Google Scholar]
  10. Durrett L. R., Ziegler M. G. A sensitive radioenzymatic assay for catechol drugs. J Neurosci Res. 1980;5(6):587–598. doi: 10.1002/jnr.490050613. [DOI] [PubMed] [Google Scholar]
  11. Eschenhagen T., Mende U., Nose M., Schmitz W., Scholz H., Warnholtz A., Wüstel J. M. Isoprenaline-induced increase in mRNA levels of inhibitory G-protein alpha-subunits in rat heart. Naunyn Schmiedebergs Arch Pharmacol. 1991 Jun;343(6):609–615. doi: 10.1007/BF00184292. [DOI] [PubMed] [Google Scholar]
  12. Feldman A. M., Cates A. E., Veazey W. B., Hershberger R. E., Bristow M. R., Baughman K. L., Baumgartner W. A., Van Dop C. Increase of the 40,000-mol wt pertussis toxin substrate (G protein) in the failing human heart. J Clin Invest. 1988 Jul;82(1):189–197. doi: 10.1172/JCI113569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feldman M. D., Copelas L., Gwathmey J. K., Phillips P., Warren S. E., Schoen F. J., Grossman W., Morgan J. P. Deficient production of cyclic AMP: pharmacologic evidence of an important cause of contractile dysfunction in patients with end-stage heart failure. Circulation. 1987 Feb;75(2):331–339. doi: 10.1161/01.cir.75.2.331. [DOI] [PubMed] [Google Scholar]
  14. Gierschik P., Milligan G., Pines M., Goldsmith P., Codina J., Klee W., Spiegel A. Use of specific antibodies to quantitate the guanine nucleotide-binding protein Go in brain. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2258–2262. doi: 10.1073/pnas.83.7.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hammond H. K., Ransnas L. A., Insel P. A. Noncoordinate regulation of cardiac Gs protein and beta-adrenergic receptors by a physiological stimulus, chronic dynamic exercise. J Clin Invest. 1988 Dec;82(6):2168–2171. doi: 10.1172/JCI113840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hammond H. K., Roth D. A., Ford C. E., Stamnas G. W., Ziegler M. G., Ennis C. Myocardial adrenergic denervation supersensitivity depends on a postreceptor mechanism not linked with increased cAMP production. Circulation. 1992 Feb;85(2):666–679. doi: 10.1161/01.cir.85.2.666. [DOI] [PubMed] [Google Scholar]
  17. Hammond H. K., Roth D. A., Insel P. A., Ford C. E., White F. C., Maisel A. S., Ziegler M. G., Bloor C. M. Myocardial beta-adrenergic receptor expression and signal transduction after chronic volume-overload hypertrophy and circulatory congestion. Circulation. 1992 Jan;85(1):269–280. doi: 10.1161/01.cir.85.1.269. [DOI] [PubMed] [Google Scholar]
  18. Hammond H. K., White F. C., Brunton L. L., Longhurst J. C. Association of decreased myocardial beta-receptors and chronotropic response to isoproterenol and exercise in pigs following chronic dynamic exercise. Circ Res. 1987 May;60(5):720–726. doi: 10.1161/01.res.60.5.720. [DOI] [PubMed] [Google Scholar]
  19. Horn E. M., Corwin S. J., Steinberg S. F., Chow Y. K., Neuberg G. W., Cannon P. J., Powers E. R., Bilezikian J. P. Reduced lymphocyte stimulatory guanine nucleotide regulatory protein and beta-adrenergic receptors in congestive heart failure and reversal with angiotensin converting enzyme inhibitor therapy. Circulation. 1988 Dec;78(6):1373–1379. doi: 10.1161/01.cir.78.6.1373. [DOI] [PubMed] [Google Scholar]
  20. Katada T., Northup J. K., Bokoch G. M., Ui M., Gilman A. G. The inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase. Subunit dissociation and guanine nucleotide-dependent hormonal inhibition. J Biol Chem. 1984 Mar 25;259(6):3578–3585. [PubMed] [Google Scholar]
  21. Katada T., Ui M. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc Natl Acad Sci U S A. 1982 May;79(10):3129–3133. doi: 10.1073/pnas.79.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Katoh Y., Komuro I., Takaku F., Yamaguchi H., Yazaki Y. Messenger RNA levels of guanine nucleotide-binding proteins are reduced in the ventricle of cardiomyopathic hamsters. Circ Res. 1990 Jul;67(1):235–239. doi: 10.1161/01.res.67.1.235. [DOI] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Linder M. E., Ewald D. A., Miller R. J., Gilman A. G. Purification and characterization of Go alpha and three types of Gi alpha after expression in Escherichia coli. J Biol Chem. 1990 May 15;265(14):8243–8251. [PubMed] [Google Scholar]
  25. Longabaugh J. P., Vatner D. E., Graham R. M., Homcy C. J. NADP improves the efficiency of cholera toxin catalyzed ADP-ribosylation in liver and heart membranes. Biochem Biophys Res Commun. 1986 May 29;137(1):328–333. doi: 10.1016/0006-291x(86)91214-3. [DOI] [PubMed] [Google Scholar]
  26. Longabaugh J. P., Vatner D. E., Vatner S. F., Homcy C. J. Decreased stimulatory guanosine triphosphate binding protein in dogs with pressure-overload left ventricular failure. J Clin Invest. 1988 Feb;81(2):420–424. doi: 10.1172/JCI113335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Luetje C. W., Tietje K. M., Christian J. L., Nathanson N. M. Differential tissue expression and developmental regulation of guanine nucleotide binding regulatory proteins and their messenger RNAs in rat heart. J Biol Chem. 1988 Sep 15;263(26):13357–13365. [PubMed] [Google Scholar]
  28. Marzo K. P., Frey M. J., Wilson J. R., Liang B. T., Manning D. R., Lanoce V., Molinoff P. B. Beta-adrenergic receptor-G protein-adenylate cyclase complex in experimental canine congestive heart failure produced by rapid ventricular pacing. Circ Res. 1991 Dec;69(6):1546–1556. doi: 10.1161/01.res.69.6.1546. [DOI] [PubMed] [Google Scholar]
  29. Nagai K., Thøgersen H. C. Generation of beta-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli. 1984 Jun 28-Jul 4Nature. 309(5971):810–812. doi: 10.1038/309810a0. [DOI] [PubMed] [Google Scholar]
  30. Negishi M., Hashimoto H., Ichikawa A. Translocation of alpha subunits of stimulatory guanine nucleotide-binding proteins through stimulation of the prostacyclin receptor in mouse mastocytoma cells. J Biol Chem. 1992 Feb 5;267(4):2364–2369. [PubMed] [Google Scholar]
  31. Neumann J., Schmitz W., Scholz H., von Meyerinck L., Döring V., Kalmar P. Increase in myocardial Gi-proteins in heart failure. Lancet. 1988 Oct 22;2(8617):936–937. doi: 10.1016/s0140-6736(88)92601-3. [DOI] [PubMed] [Google Scholar]
  32. Oduro K. A. Glycopyrrolate methobromide: 2. comparison with atropine sulphate in anaesthesia. Can Anaesth Soc J. 1975 Jul;22(4):466–473. doi: 10.1007/BF03004861. [DOI] [PubMed] [Google Scholar]
  33. Riegger G. A., Liebau G., Holzschuh M., Witkowski D., Steilner H., Kochsiek K. Role of the renin-angiotensin system in the development of congestive heart failure in the dog as assessed by chronic converting-enzyme blockade. Am J Cardiol. 1984 Feb 1;53(4):614–618. doi: 10.1016/0002-9149(84)90040-7. [DOI] [PubMed] [Google Scholar]
  34. Roth D. A., Urasawa K., Leiber D., Insel P. A., Hammond H. K. A substantial proportion of cardiac Gs is not associated with the plasma membrane. FEBS Lett. 1992 Jan 13;296(1):46–50. doi: 10.1016/0014-5793(92)80400-b. [DOI] [PubMed] [Google Scholar]
  35. Salomon Y., Londos C., Rodbell M. A highly sensitive adenylate cyclase assay. Anal Biochem. 1974 Apr;58(2):541–548. doi: 10.1016/0003-2697(74)90222-x. [DOI] [PubMed] [Google Scholar]
  36. Schuler G., Hambrecht R., Schlierf G., Niebauer J., Hauer K., Neumann J., Hoberg E., Drinkmann A., Bacher F., Grunze M. Regular physical exercise and low-fat diet. Effects on progression of coronary artery disease. Circulation. 1992 Jul;86(1):1–11. doi: 10.1161/01.cir.86.1.1. [DOI] [PubMed] [Google Scholar]
  37. Spinale F. G., Zellner J. L., Tomita M., Crawford F. A., Zile M. R. Relation between ventricular and myocyte remodeling with the development and regression of supraventricular tachycardia-induced cardiomyopathy. Circ Res. 1991 Oct;69(4):1058–1067. doi: 10.1161/01.res.69.4.1058. [DOI] [PubMed] [Google Scholar]
  38. Spinale F. G., Zellner J. L., Tomita M., Tempel G. E., Crawford F. A., Zile M. R. Tachycardia-induced cardiomyopathy: effects on blood flow and capillary structure. Am J Physiol. 1991 Jul;261(1 Pt 2):H140–H148. doi: 10.1152/ajpheart.1991.261.1.H140. [DOI] [PubMed] [Google Scholar]
  39. Sternweis P. C., Northup J. K., Smigel M. D., Gilman A. G. The regulatory component of adenylate cyclase. Purification and properties. J Biol Chem. 1981 Nov 25;256(22):11517–11526. [PubMed] [Google Scholar]
  40. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wilson J. R., Douglas P., Hickey W. F., Lanoce V., Ferraro N., Muhammad A., Reichek N. Experimental congestive heart failure produced by rapid ventricular pacing in the dog: cardiac effects. Circulation. 1987 Apr;75(4):857–867. doi: 10.1161/01.cir.75.4.857. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES