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Modification of contractile proteins by oxygen free radicals in rat heart

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Abstract

This study was undertaken to investigate the effects of oxygen free radicals on myofibrillar creatine kinase activity. Isolated rat heart myofibrils were incubated with xanthine+xanthine oxidase (a superoxide anion radical-generating system) or hydrogen peroxide and assayed for creatine kinase activity. To clarify the involvement of changes in sulfhydryl groups in causing alterations in myofibrillar creatine kinase activity, 1) effects of N-ethylmaleimide (sulfhydryl groups reagent) on myofibrillar creatine kinase activity, 2) effect of oxygen free radicals on myofibrillar sulfhydryl groups content, and 3) protective effects of dithiothreitol (sulfhydryl groups-reducing agent) on the changes in myofibrillar creatine kinase activity due to oxygen free radicals were also studied. Xanthine+xanthine oxidase inhibited creatine kinase activity both in a time-and a concentration-dependent manner. Superoxide dismutase (SOD) showed a protective effect on the depression in creatine kinase activity caused by xanthine+xanthine oxidase. Hydrogen peroxide inhibited creatine kinase activity in a concentration-dependent manner; this inhibition was prevented by the addition of catalase. N-ethylmaleimide reduced creatine kinase activity in a dose-dependent manner. The content of myofibrillar sulfhydryl groups was decreased by xanthine+xanthine oxidase; this reduction was protected by SOD. Furthermore, the depression in myofibrillar creatine kinase activity by xanthine+xanthine oxidase was protected by the addition of dithiothreitol. Oxygen free radicals may inhibit myofibrillar creatine kinase activity by modifying sulfhydryl groups in the enzyme protein. The reduction of myofibrillar creatine kinase activity may lead to a disturbance of energy utilization in the heart and may contribute to cardiac dysfunction due to oxygen free radicals.

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References

  1. Feeman BA, Crapo JD: Biology of disease: Free radicals and tissue injury. Lab Invest 47: 412–426, 1982

    Google Scholar 

  2. Halliwell B, Grootreld M: The measurement of free radicals in humans. FEBS Lett 213: 9–14, 1986

    Google Scholar 

  3. Gupta M, Singal PK: Oxygen radical injury in the presence of desferal, a specific iron-chelating agent. Biochem Pharmacol 36: 3774–3777, 1987

    Google Scholar 

  4. Przyklenk K, Whittaker P, Kloner RA: Direct evidence that oxygen free radicals cause contractile dysfunctionin vivo. Circulation 78 (Suppl II): II-264, 1988

    Google Scholar 

  5. Saks VA, Ventura-Clapier R, Huchua ZA, Preobrazhensky AN, Emelin IV: Creatine kinase in regulation of heart function and metabolism. I. Further evidence for compartmentation of adenine nucleotides in cardiac myofibrillar and sarcolemmal coupled ATPase-creatine kinase systems. Biochem Biophys Acta 803: 254–264, 1984

    Google Scholar 

  6. Wallimann T, Schlosser T, Eppenberger HM: Function of M-line-bound creatine kinase in myofibrillar ATP regenerator at the receiving end of the phosphorylcreatine shuttle in muscle. J Biol Chem 259: 5238–5246, 1984

    Google Scholar 

  7. Bessman SP, Yang WCT, Geiger PJ, Erickson-Viitanen S: Intimate coupling of creatine phosphokinase and myofibrillar adenosine triphosphatase. Biochem Biophys Res Commun 96: 1414–1420, 1980

    Google Scholar 

  8. Yagi K, Mase R: Coupled reaction of creatine kinase and myosin A-adenosine triphosphatase. J Biol Chem 259: 5238–5246, 1984

    Google Scholar 

  9. Fossel ET, Hoefeler H: Complete inhibition of creatine kinase in isolated perfused rat hearts. Am J Physiol 252 (Endocrinol Metab 15): E124–E130, 1987

    Google Scholar 

  10. Kaneko M, Beamish RE, Dhalla NS: Depression of heart sarcolemmal Ca2+-pump activity by oxygen free radicals. Am J Physiol 256 (Heart Circ Physiol 25): H368–H374, 1989

    Google Scholar 

  11. Kaneko M, Elimban V, Dhalla NS: Mechanism for depression of heart sarcolemmal Ca2+-pump by oxygen free radicals. Am J Physiol 257 (Heart Circ Physiol 26): H804–H811, 1989

    Google Scholar 

  12. Kaneko M, Lee SL, Wolf CM, Dhalla NS: Reduction of calcium channel antagonist binding sites by oxygen free radicals in rat heart. J Mol Cell Cardiol 21: 935–943, 1989

    Google Scholar 

  13. Kaneko M, Chapman DC, Ganguly PK, Beamish RE, Dhalla NS: Modification of cardiac adrenergic receptors by oxygen free radicals. Am J Physiol 260 (Heart Cric Physiol 29): H821–H826, 1991

    Google Scholar 

  14. Okabe E, Hess ML, Oyama M, Ito H: Characterization of free radical-mediated damage of canine cardiac sarcoplasmic reticulum. Arch Biochem Biophys 225: 164–177, 1983

    Google Scholar 

  15. Otani H, Tanaka H, Inoue T, Umemoto M, Omoto K, Tanaka K, Sato T, Osako T, Masuda A, Nonoyama A, Kagawa T.In vivo study on contribution of oxidative metabolism of isolated rabbit heart mitochondria to myocardial reperfusion injury. Circ Res 55: 168–175, 1984

    Google Scholar 

  16. Solaro RJ, Pang DC, Briggs FN: The purification of cardiac myofibrils with Triton X-100. Biochem Biophys Acta 245: 259–262, 1971

    Google Scholar 

  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  18. Hess JW, MacDonald RP, Natho GJW, Murdock KJ: Serum creatine phosphokinase: Evaluation of a commercial spectrophotometric method. Clin Chem 13: 994–1005, 1967

    Google Scholar 

  19. Schoot BM, Schoots AFM, Depont JJHHM, Schuurmans-Stekhoven FMAH, Bonting SL: Studies on (Na++K+) activated ATPase. XLI: Effects of N-ethylmaleimide on overall and partial reactions. Biochim Biophys Acta 483: 181–192, 1977

    Google Scholar 

  20. Yamada S, Ikemoto N: Distinction of thiols involved in the specific reaction steps of the Ca2+-ATPase of the sarcoplasmic reticulum. J Biol Chem 253: 6801–6807, 1987

    Google Scholar 

  21. Trundle D, Cunningham LW: Iodine oxidation of the sulfhydryl groups of creatine kinase. Biochemistry 8: 1919–1925, 1969

    Google Scholar 

  22. Bolli R, Jeroudi MO, Patel BS, DuBose CM, Lai EK, Robert R, McCay PB: Direct evidence that oxygen-drived free radicals contribute to postischemic myocardial dysfunction in intact dog. Proc Natl Acad Sci USA 86: 4695–4699, 1989

    Google Scholar 

  23. Zweier JL, Flaherty JT, Weisfedt ML: Direct measurement of free radical generation following reperfusion of ischemic myocardium. Proc Natl Acad Sci USA 84: 1404–1407, 1987

    Google Scholar 

  24. Greenfield RA, Swain JL: Disruption of myofibrillar energy use: Dual mechanism that may contribute to postischemic dysfunction in stunned myocardium. Circ Res 60: 283–289, 1987

    Google Scholar 

  25. Vial C, Font B, Goldschmidt P, Gantheron DC: Dissociation and reassociation of creatine kinase with heart mitochondria: pH and phosphate dependence. Biochem Res Commun 88: 1352–1359, 1979

    Google Scholar 

  26. Bittle JA, Weisfeldt ML, Jacobus WE: Creatine kinase of heart mitochondria: Progressive loss of energy activity duringin vivo ischemia and its correlation to depressed myocardial function. J Biol Chem 260: 208–214, 1985

    Google Scholar 

  27. Suzuki S, Kaneko M, Chapman DC, Dhalla NS: Alterations in cardiac contractile proteins due to oxygen free radicals. Biochem Biophys Acta 1074: 95–100, 1991

    Google Scholar 

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Authors and Affiliations

  1. The Third Department of Internal Medicine, Hamamatsu University School of Medicine, 3600 Handa-cho, 431-31, Hamamatsu, Japan

    Masanori Kaneko, Hisamichi Masuda, Hideki Suzuki, Yuji Matsumoto, Akira Kobayashi & Noboru Yamazaki

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  1. Masanori Kaneko
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  2. Hisamichi Masuda
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  3. Hideki Suzuki
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  4. Yuji Matsumoto
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  5. Akira Kobayashi
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  6. Noboru Yamazaki
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Kaneko, M., Masuda, H., Suzuki, H. et al. Modification of contractile proteins by oxygen free radicals in rat heart. Mol Cell Biochem 125, 163–169 (1993). https://doi.org/10.1007/BF00936445

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  • DOI: https://doi.org/10.1007/BF00936445

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