Advertisement

Molecular and Cellular Biochemistry

, Volume 269, Issue 1, pp 49–57 | Cite as

Reduced inhibitor 1 and 2 activity is associated with increased protein phosphatase type 1 activity in left ventricular myocardium of one-kidney, one-clip hypertensive rats

  • Ramesh C. GuptaEmail author
  • Sudhish Mishra
  • Xiao-Ping Yang
  • Hani N. Sabbah
Article

Abstract

In failing hearts, although protein phosphatase type 1 (PP1) activity has increased, information about the regulation and status of PP1 inhibitor-1 (INH-1) and inhibitor-2 (INH-2) is limited. In this study, we examined activity and protein expression of PP1, INH-1 and INH-2 and phosphorylation of sarcoplasmic reticulum (SR) phospholamban (PLB), a substrate of PP1 and modulator of SR Ca2+-ATPase activity, in failing and non-failing hearts. These studies were performed in LV myocardium of seven rats with chronic renal hypertension produced by Goldblatt’s one-kidney, one-clip procedure and seven age-matched sham-operated normal controls (CTR). Eight weeks after surgery, LV ejection fraction, LV hypertrophy, and pulmonary congestion were determined in all rats. PP1 activity (nmol 32P/min/mg non-collagen protein) was assessed in LV homogenates using 32P-labeled phosphorylase a as substrate. INH-1 and INH-2 activity was determined in the immunoprecipitate of LV homogenates and expressed as percentage inhibitory activity. Using a specific antibody, LV tissue levels of PP1C and calsequestrin (CSQ), a SR calcium binding protein, which is not altered in failing hearts, were also determined. Further, total and phosphorylated PLB, INH-1 and INH-2 protein levels were determined in the LV homogenate and phosphoprotein-enriched fraction, respectively. The band density of each protein was quantified in densitometric units and normalized to CSQ. Results: rats with chronic renal hypertension exhibited significantly reduced LV ejection fraction and increased LV hypertrophy and pulmonary congestion, characteristics of chronic heart failure (CHF). We found that compared to CTR, (1) both INH-1 (10.2 ± 2 versus 57.5 ± 1; p<0.05) and INH-2 activity (3.8 ± 0.4 versus 36.2 ± 4; p<0.05) were reduced, (2) total and phosphorylated PLB amount reduced, (3) protein level of phosphorylated INH-1 was reduced (2.32 ± 0.1 versus 0.73 ± 0.04; p<0.05) whereas that of phosphorylated INH-2 increased (3.05 ± 0.3 versus 1.42 ± 0.1; p<0.05), and (4) PP1 activity was increased approximately 2.6-fold in rats with CHF (1.59 ± 0.05 versus 0.61 ± 0.01; p<0.05) while protein level of the catalytic subunit of PP1 (PP1C) increased 3.85-fold (0.77 ± 0.05 versus 0.20 ± 0.02; p<0.05). These results suggest that reduced inhibitory INH-1 and INH-2 activity, increased PP1C protein level, and reduced PLB phosphorylation are associated with increased PP1 activity in failing hearts. (Mol Cell Biochem 269: 49–57, 2005)

Keywords

heart failure protein phosphatases inhibitor-1 inhibitor-2 phospholamban protein phosphorylation 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Pathophysiology of heart failure. Identifying targets for pharmacotherapy. Med Clin North Am 87: 303–316, 2003PubMedGoogle Scholar
  2. 2.
    Petrashevskaya NN, Koch SE, Bodi I, Schwartz A: Calcium cycling, historic overview and perspectives. Role for autonomic nervous system regulation. J Mol Cell Cardiol 34: 885–896, 2002Google Scholar
  3. 3.
    Hersiz S, Neumann J: Effects of serine/threonine protein phosphatases on ion channels in excitable membranes. Physiol Rev 80: 173–210, 2000PubMedGoogle Scholar
  4. 4.
    Haq S, Choukroun G, Lim H, Tymitz KM, del Monte F, Gwathmey J, Grazette L, Michael A, Hajjar R, Force T, Molkentin JD: Differential activation of signal transduction pathways in human hearts with hypertrophy versus advanced heart failure. Circulation 103: 670–677, 2001PubMedGoogle Scholar
  5. 5.
    Molkentin JD, Lu JR, Antos CL, Markham B, Richardson J, Robbins J, Grant SR, Olson EN: A calcineurin-dependent transcriptional pathway for cardiac hypertrophy. Cell 93: 215–228, 1998CrossRefPubMedGoogle Scholar
  6. 6.
    Oie E, Bjornerheim R, Clausen OP, Attramadal H: Cyclosporin A inhibits cardiac hypertrophy and enhances cardiac dysfunction during postinfarction failure in rats. Am J Physiol Heart Circ Physiol 278: H2115–H2123, 2000Google Scholar
  7. 7.
    Chu G, Carr AN, Young KB, Lester JW, Yatani A, Sanbe A, Colbert MC, Schwartz SM, Frank KF, Lampe PD, Robbins J, Molkentin JD, Kranias EG: Enhanced myocyte contractility and Ca2+handling in a calcineurin transgenic model of heart failure. Cardiovasc Res 54: 105–116, 2002Google Scholar
  8. 8.
    Solaro RJ: Protein phosphorylation and cardiac myofilaments. In: RJ Solaro ed. Protein Phosphorylation in Heart Muscle, CRC Press, Boca Raton, FL, 1986, pp 129–156Google Scholar
  9. 9.
    Gupta RC, Neumann J, Durant P, Watanabe AM: Comparison of adenosine and muscarinic receptor-mediated effects of protein phosphatase inhibitor activity in the heart. J Pharmacol Exp Ther 266: 16–22, 1993Google Scholar
  10. 10.
    Gupta RC, Neumann J, Watanabe AM, Lesch M, Sabbah HN: Evidence for presence and hormonal regulation of protein phosphatase inhibitor-1 in ventricular cardiomyocytes. Am J Physiol Heart Circ Physiol 270: H1159–H1164, 1996Google Scholar
  11. 11.
    Neumann J, Gupta RC, Schmitz W, Scholz H, Nairn AC, Watanabe AM: Evidence for isoproterenol-induced phosphorylation of protein phosphatase inhibitor-1 in the intact heart. Circ Res 69: 1450–1457, 1991Google Scholar
  12. 12.
    Gupta RC, Neumann J, Watanabe AM, Sabbah HN: Inhibition of type 1 protein phosphatase activity by activation of β-adrenoceptors in ventricular myocardium. Biochem Pharmacol 63: 1069–1076, 2002Google Scholar
  13. 13.
    Wang QM, Guan KL, Roach PJ, DePaoli-Roach AA: Phosphorylation and activation of the ATP-Mg2+-dependent protein phosphatase by the mitogen-activated protein kinase. J Biol Chem 270: 18352–18358, 1995Google Scholar
  14. 14.
    Neumann J, Eschenhagen T, Jones LR, Linck B, Schmitz W, Scholz H, Zimmermann N: Increased expression of cardiac phosphatases in patients with end-stage heart failure. J Mol Cell Cardiol 29: 265–272, 1997Google Scholar
  15. 15.
    Huang B, Wang S, Qin D, Boutjdir M, El-Sherif N: Diminished basal phosphorylation level of phospholamban in the postinfarction remodeled rat ventricle: Role of β-adrenergic pathway, Gi protein, phosphodiesterase, and phosphatases. Circ Res 85: 848–855, 1999Google Scholar
  16. 16.
    Nettican T, Temsah RM, Kawabata K, Dhalla NS: Sarcoplasmic reticulum Ca2+calmodulin-dependent protein kinase is altered in heart failure. Circ Res 86: 596–605, 2000Google Scholar
  17. 17.
    Mishra S, Gupta RC, Tiwari N, Sharov VG, Sabbah HN: Molecular mechanisms of reduced sarcoplasmic reticulum Ca2+uptake in human failing left ventricular myocardium. J Heart Lung Transplant 21: 366–373, 2002Google Scholar
  18. 18.
    El-Armouche A, Pamminger T, Ditz D, Zolk O, Eschenhagen T: Decreased protein and phosphorylation level of the protein phosphatase inhibitor-1 in failing hearts. Cardiovasc Res 61: 87–93, 2004Google Scholar
  19. 19.
    Carr AN, Schmidt AG, Suzuki Y, Monte FD, Sato Y, Lanner C, Breeden K, Jing SL, Allen PB, Greengard P, Yatani A, Hoit BD, Grupp IL, Hajjar RJ, DePaoli-Roach AA, Kranias EG: Type-1 phosphatase, a negative regulator of cardiac function. Mol Cell Biol 22: 4124–4135, 2002Google Scholar
  20. 20.
    Braz JC, Gregory K, Pathak A, Zhao W, Sahin B, Klevitsky R, Kimball TF, Lorenz JN, Nairn AC, Liggett SB, Bodi I, Wang S, Schwartz A, Lakatta EG, De Paoli-Roach AA, Robbins J, Hewett TE, Bibb JA, Westfall MV, Kranias EG, Molkentin JD: PKC-a regulates cardiac contractility and propensity toward heart failure. Nat Med 10: 248–254, 2004Google Scholar
  21. 21.
    Gupta RC, Mishra S, Rastogi S, Imai M, Habib O, Sabbah HN: Cardiac SR-coupled PP1 activity and expression are increased and inhibitor-1 expression is decreased in failing hearts. Am J Physiol Heart Circ Physiol 285: H2373–H2381, 2003Google Scholar
  22. 22.
    Gupta RC, Yang XP, Mishra S, Sabbah HN: Assessment of sarcoplasmic reticulum Ca2+-uptake during the development of left ventricular hypertrophy. Biochem Pharmacol 65: 933–939, 2003Google Scholar
  23. 23.
    Kirchhefer U, Schmitz W, Scholz H, Neumann J: Activity of cAMP-dependent protein kinase and Ca2+calmodulin-dependent protein kinase in failing and nonfailing human hearts. Cardiovasc Res 42: 254–261, 1999Google Scholar
  24. 24.
    Liu YH, Yang XP, Nass O, Sabbah HN, Peterson E, Carretereo OA: Chronic heart failure induced by coronary artery ligation in Lewis inbred rats. Am J Physiol Heart Circ Physiol 272: H722–H727, 1997Google Scholar
  25. 25.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1951Google Scholar
  26. 26.
    Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976CrossRefPubMedGoogle Scholar
  27. 27.
    Gupta RC, Shimoyama H, Tanimura M, Nair R, Lesch M, Sabbah HN: SR Ca2+-ATPase activity and expression in ventricular myocardium of dogs with heart failure. Am J Physiol 273: H12–H18, 1997Google Scholar
  28. 28.
    Gupta RC, Mishra S, Mishima T, Goldstein S, Sabbah HN: Reduced sarcoplasmic reticulum Ca2+-uptake and expression of phospholamban in left ventricular myocardium of dogs with heart failure. J Mol Cell Cardiol 31: 1381–1389, 1999Google Scholar
  29. 29.
    Sventek P, Turgeon A, Garcia R, Schiffrin EL: Vascular and cardiac overexpression of endothelin-1 gene in one-kidney, one clip Goldblatt hypertensive rats but only in the late phase of two-kidney one clip Goldblatt hypertension. J Hypertens 14: 57–64, 1996Google Scholar
  30. 30.
    Ge C, Garcia R, Anand-Srivastava MB: Altered expression of Gi-protein and adenylyl cyclase activity in hearts from one kidney one clip hypertensive rats: Effect of captopril. J Hypertens 17: 1617–1626, 1999Google Scholar
  31. 31.
    Sabbah HN, Sharov VG, Gupta RC, Mishra S, Rastogi S, Undrovinas AI, Chaudhry PA, Todor A, Mishima T, Tanhehco EJ, Suzuki G: Reversal of chronic molecular and cellular abnormalities due to heart failure by passive mechanical ventricular containment. Circ Res 93: 1020–1022, 2003Google Scholar
  32. 32.
    Kaiser RA, Bueno OF, Lips DJ, Doevendans PA, Jones F, Kimball TF, Molkentin JD: Targeted inhibition of p38 MAPK antagonizes cardiac injury and cell death following ischemia-reperfusion in vivo. J Biol Chem 279: 15524–15530, 2004Google Scholar
  33. 33.
    Everett AD, Stoops TD, Nairn AC, Brautigan D: Angiotensin II regulates phosphorylation of translation elongation factor-2 in cardiac myocytes. Am J Physiol Heart Circ Physiol 281: H161–H167, 2001Google Scholar
  34. 34.
    Chen X, Piacentino V 3rd, Furukawa S, Goldman B, Margulies KB, Houser SR: L-type Ca2+channel density and regulation are altered in failing human ventricular myocytes and recover after support with mechanical assist devices. Circ Res 91: 517–524, 2002Google Scholar
  35. 35.
    Wei SK, Ruknudin A, Hanlon SU, McCurley JM, Schulze DH, Haigney MC: Protein kinase A hyperphosphorylation increases basal current but decreases beta-adrenergic responsiveness of the sarcolemmal Na+-Ca2+exchanger in failing pig myocytes. Circ Res. 92: 897–903, 2003Google Scholar
  36. 36.
    Reiken S, Gaburjakova M, Guatimosim S, Gomez AM, D’Armiento J, Burkhoff D, Wang J, Vassort G, Lederer WJ, Marks AR: Protein kinase A phosphorylation of the cardiac calcium release channel ryanodine receptor in normal and failing hearts. Role of phosphatases and responses to isoproterenol. J Biol Chem 278: 444–453, 2003Google Scholar
  37. 37.
    Horiuchi M, Hayashida W, Kambe T, Yamada T, Dzau VJ: Angiotensin type 2 receptor dephosphorylates Bcl-2 by activating mitogen-activated protein kinase phosphatase-1 and induces apoptosis. J Biol Chem 272: 19022–19026, 1997Google Scholar
  38. 38.
    Mishra S, Sabbah HN, Jain JC, Gupta RC: Reduced Ca2+-calmodulin-dependent protein kinase activity and expression in LV myocardium of dogs with heart failure. Am J Physiol Heart Circ Physiol 284: H876–H883, 2003Google Scholar
  39. 39.
    El-Armouche A, Rau T, Zolk O, Ditz D, Pamminger T, Zimmermann WH, Jackel E, Harding SE, Boknik P, Neumann J, Eschenhagen T: Evidence for protein phosphatase inhibitor-1 playing an amplifier role in beta-adrenergic signaling in cardiac myocytes. FASEB J 17: 437–439, 2003Google Scholar
  40. 40.
    Williams JP, Jo H, Hunnicut RE, Brautigan DL, McDonald JM: Tyrosine phosphorylation of phosphatase inhibitor 2. J Cell Biochem 57: 415–422, 1995Google Scholar
  41. 41.
    Takeishi Y, Huang Q, Abe JI, Che W, Lee JW, Kawakatsu H, Hoit BD, Berk BC, Walsh RA: Activation of mitogen-activated protein kinases and p90 ribosomal S6 kinase in failing human hearts with dilated cardiomyopathy. Cardiovasc Res 53: 131–137, 2002Google Scholar

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Ramesh C. Gupta
    • 1
    Email author
  • Sudhish Mishra
    • 1
  • Xiao-Ping Yang
    • 2
  • Hani N. Sabbah
    • 1
  1. 1.Cardiovascular Medicine, Department of MedicineHenry Ford Heart and Vascular Institute, Henry Ford Health SystemDetroitUSA
  2. 2.Hypertension and Vascular Research Division, Department of MedicineHenry Ford Heart and Vascular Institute, Henry Ford Health SystemDetroitUSA

Personalised recommendations