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Na+/Ca2+ exchange of isolated sarcolemmal membrane: effects of insulin, oxidants and insulin deficiency

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Abstract

In this study we prepared sarcolemmal fractions from bovine and rat hearts; their Na+K+ ATPase activities, measured in the presence of saponin to unmask latent Na+K+ ATPase, were 59.4 and 48.8 µ mol Pi/mg protein · h, respectively. The rate of Na+dependent Ca2+ uptake was linear for the first 10 s and a plateau was reached in 3 min. Oxidation by free radical generation either with H2O2, FeSO4 plus DTT or xanthine oxidase plus hypoxanthine stimulated Na+/Ca2+ exchange in a time-dependent manner. The stimulation was abolished by deferoxamine or o-phenanthroline. By contrast, oxidation by HOCI inhibited Na+/Ca2+ exchange in proportion to its concentration, and this inhibition was antagonized by DTT. DTT alone had no effect on the exchange. Insulin stimulated Na+/Ca2+ exchange, its maximal effect was attained after 30min incubation with 100 µ units/ml. N-ethylmaleimide inhibited the exchange both in the presence and in the absence of insulin. Sarcolemmal fractions prepared from hearts of alloxan-treated, acutely diabetic rats showed a significant decrease in Na+/Ca2+ exchange. Addition of insulin in vitro significantly stimulated Na+/Ca2+ exchange of both diabetic and control groups. The results indicate that sarcolemmal Na+/Ca2+ exchange function is modulated by oxidation-reduction states and by the presence of insulin.

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References

  1. Takahashi K, Kako KJ: Ischemia-induced changes in sarcolemmal Na+K+ ATPase, K+ pNPPase, sialic acid and phospholipid in the dog and effects of the Nisoldipine and chlorpromazine treatment. Biochem Med 31:271–286, 1984

    Google Scholar 

  2. Kako KJ, Kato M, Matsuoka T, Mustapha A: The depression of membrane-bound Na+K+ ATPase activity induced by free radicals and by ischemia of the kidney. Am J Physiol 254:C330-C337 1988

    Google Scholar 

  3. Kato M, Kako KJ: Effect of N-(2-mercaptopropionyl)glycine on ischemic-reperfused dog kidney in vivo and membrane preparation in vitro. Mol Cell Biochem 78:151–159 (1987)

    Google Scholar 

  4. Dhalla NS, Pierce GN, Panagia V, Signal PK, Beamish RE: Calcium movements in relation to heart function. Basic Res Cardiol 77:117–139, 1982

    Google Scholar 

  5. Dhalla NS, Pierce GN, Innes IR, Beamish RE: Pathogenesis of cardiac dysfunction in diabetes mellitus. Can J Cardiol 1:263–281, 1985

    Google Scholar 

  6. Ganguly PK, Pierce GN, Dhalla KS, Dballa NS: Defective sarcoplasmic reticular calcium transport in diabetic cardiomyopathy. Am J Physiol 244:E528–535, 1983

    Google Scholar 

  7. Pierce GN, Kutryk MJB, Dhalla NS: Alterations in Ca binding by and composition of the cardiac sarcolemmal membrane in chronic diabetes. Proc Nat Acad Sci US 80:5412–5516, 1983

    Google Scholar 

  8. Makino N, Dhalla KS, Elimban V, Dhalla NS: Sarcolemmal Ca transport in streptozotocin-induced diabetic cardiomyopathy in rats. Am J Physiol 253:E202–207, 1987

    Google Scholar 

  9. Heyliger CE, Prakash A, McNeil JH: Alterations in cardiac sarcolemmal Ca pump activity during diabetes mellitus. Am J Physiol 252:H540–544, 1987

    Google Scholar 

  10. Pierce GN, Dhalla NS: Cardiac myofibrillar ATPase activity in diabetic rats. J Mol Cell Cardiol 13:1063–1069, 1981

    Google Scholar 

  11. Ganguly PK, Rice KM, Panagia V, Dhalla NS: Sarcolemmal phosphatidylethanolamine N-methylation in diabetic cardiomyopathy. Circ Res 55:504–512, 1984

    Google Scholar 

  12. Czech MP, Lawrence Jr JC, Lynn WS: Evidence for the involvement of sulfhydryl oxidation in the regulation of fat cell bexose transport by insulin. Proc Nat Acad Sci USA 71:4173–4177, 1974

    Google Scholar 

  13. Ramasarma T: Generation of H2O2 in biomembranes. Biochim Biophys Acta 694:69–93, 1982

    Google Scholar 

  14. Hayes GR, Lockwood DH: Role of insulin receptor phosphorylation in the the insulinomimetic effects of hydrogen peroxide. Proc Nat Acad Sic USA 84:8115–8119, 1987

    Google Scholar 

  15. Kozka IJ, Gould MK: Inhibitory effects of Nethylmaleimide on insulin- and oxidant-stimulated sugar transport and on 125I-labelled insulin binding by rat soleus muscle. Biochim Biophys Acta 797:212–220, 1984

    Google Scholar 

  16. Pershadsingh HA, Shade DL, Delfert DM, McDonald JM: Chelation of intracellular calcium blocks insulin arction in the adipocyte. Proc Nat Acad Sci USA 84:1025–1029, 1987

    Google Scholar 

  17. Takasu N, Yamada T, Shimizu Y: Generation of H202 is regulated by cytoplasmic free calcium in cultured porcine thyroid cells. Biochem Biohphys Res Comm 148:1527–1532, 1987

    Google Scholar 

  18. Orrenius S and Bellomo G: Toxicological implications of perturbation of Ca2+ homeostasis in hepatocytes. Calcium Cell Funct. 4:185–208, 1986

    Google Scholar 

  19. Carafoli E, Longoni S: The plasma membrane in the control of the signaling function of calcium. In IJ Mandel and DC Eaton (eds). Cell Calcium and the Control of Membrane Transport, Rockefeller Univ. Press, New York, 21–30, 1986

    Google Scholar 

  20. Parker JC: Diamide stimulates calcium-sodium exchange in dog red blood cells. Am J Physiol 253:C580–587, 1987

    Google Scholar 

  21. Carpentier JL, Gordon P, Robert A, Orci L: Internalization of polypeptide hormones and receptor recycling. Experientia 42:734–744, 1986

    Google Scholar 

  22. Mansier P, Charlemagne D, Rossi B, Preteseille M, Swynghedauw B, Lelievre L: Isolation of impermeable inside-out vesicles from an enriched sarcolemma fraction of rat heart. J Biol Chem 258:6628–6635, 1983

    Google Scholar 

  23. Lukens FDW: Alloxan diabetes. Physiol Rev 28:304–330, 1948

    Google Scholar 

  24. Rerup CC: Drugs producing diabetes through damage of the insulin secreting cells. Pharmacol Rev 22:485–518, 1970

    Google Scholar 

  25. Bell RH, Hye RJ: Animal models of diabetes mellitus. Physiology and pathology. J Surg Res 35:433–460, 1983

    Google Scholar 

  26. Philipson KID: Methods for measuring sodium-calcium exchange in cardiac sarcolemmal vesicles. In: NS Dhalla (ed) Methods in Studying Cardiac Membranes. 1984, pp 147–155

  27. Wharton DC, Tzagoloff A: Cytochrome oxidase from beef heart mitochondria. Methods Enzymol. 10:245–250, 1967

    Google Scholar 

  28. Aronson Jr NN, Touster O: Isolation of rat liver plasma membrane fragments in isotonic sucrose. Methods Enzymol. 31:93–102, 1974

    Google Scholar 

  29. Hidalgo C, Parra C, Rioquelme G, Jaimovich E: Transverse tubules from frog skeletal muscle. Purification and properties of vesicles sealed with the inside-out orientation. Biochim Biophys Acta 855:79–88, 1986

    Google Scholar 

  30. Takahashi K, Kako KJ: The effect of a calcium channel antagonist, Nisoldipine, on the ischemia-induced change of canine sarcolemmal membrane. Basic Res Cardiol 78:326–337, 1983

    Google Scholar 

  31. Jones LR, Maddock SW, Besch Jr HR: Unmasking effect of alamethicin on the Na+K+ ATPase, beta adrenergic receptorcoupled adenylate cyclase and cAMP-depdendent protein kinase activities of cardiac sarcolemmal vesicles. J Biol Chem 255:9971–9980, 1980

    Google Scholar 

  32. Peterson CL: A simplification of the protein assay method of Lowry et al which is more generally applicable. Anal Biochem 83:346–356, 1977

    CAS  PubMed  Google Scholar 

  33. Kato M, Kako KJ: Orientation of vesicles isolated from basolateral membranes of renal cortex. Mol Cell Biochem 78:9–16, 1987

    Google Scholar 

  34. Reeves JP, Bailey CA, Hale CC: Redox modification of sodium-calcium exchange activity in cardiac sarcolemmal vesicles. J Biol Chem 261:4948–4955, 1986

    Google Scholar 

  35. Aust SD, Morehouse LA, Thomas CE: Role of metals in oxygen radical reactions. J Free Radicals Biol Med 1:3–25, 1985

    Google Scholar 

  36. Halliwell B, Gutteridge JMC: The importance of free radicals and catalytic: metal ions in human diseases. Mol Asp Med 8:89–193, 1985

    Google Scholar 

  37. Kako KJ: Membrane damage caused by lipid peroxidation in myocardial ischemia. Jikei Med J 32:609–639, 1985

    Google Scholar 

  38. Matsuoka T, Kato M, Kako KJ: Effects of hydrogen peroxide and hypochlorite on Na+K+ ATPase. (MS submitted) (Abstr) J Mol Cell Cardiol 19: Suppl IV-78, 1987

  39. Weiss SJ, Klein R, Slivka A, Wei M: Chlorination of taurine by human neutrophils. Evidence for hyochlorous acid generation. J Clin Invest 70:598–607, 1982

    Google Scholar 

  40. Gupta MP, Makino N, Khatter K, Dhalla NS: Stimulation of Na+Ca2+ exchange in heart sarcolemma by insulin. Life Sci. 39:1077–1083, 1986

    Google Scholar 

  41. Goldstein S, Czapski G: The role and mechanism of metal ions and their complexes in enhancing damage in biological systems or in protecting these systems from the toxicity of O2. J Free Radicals Biol Med 2:3–11, 1986

    Google Scholar 

  42. Levine RL: Covalent modification of proteins by mixed function oxidation. Curr Top Cell Regul 27:305–316, 1985

    Google Scholar 

  43. Yip CC, Moule ML: Structure of the insulin receptor of rat adipocytes. Diabetes 32:760–767, 1983

    Google Scholar 

  44. Winterbourne CC: Comparative reactivities of various biological compounds with myeloperoxidase-hydrogen peroxide-chloride, and similarity of the oxidant to hypochlorite. Biochim Biophys Acta 840:204–210, 1985

    Google Scholar 

  45. Kako KJ, Yanagishita T, Kato M, Kaminishi T, Matsuoka T: Mechanisms of oxidant-induced perturbation of calcium homeostasis in heart cells. In Medical and Biochemical Aspects of Free Radicals, Yoshikawa T (ed), Elsevier Publ. (in press) 1988

  46. Rivett AJ, Roseman JE, Oliver CN, Levine RL, Stadtman ER: Covalent modification of proteins by mixed-function oxidation. Recognition by intracellular proteases. In Intracellular Protein Catabolism, ed EA Khairallah, JS Bond, JWC Bird, AR Liss, New York, 317–328, 1985

    Google Scholar 

  47. Davies KJA, Goldberg AL: Oxygen radicals stimulate intracellular proteolysis and lipid peroxidation by independent mechanisms in erythrocytes. J Biol Chem 262:8220–8226, 1987

    Google Scholar 

  48. 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. 1978

    Google Scholar 

  49. Saito K, Yamashita T, Kubota I, Kawakita M: Reactive sulfhydryl groups of sarcoplasmic reticulum ATPase. J. Biochem. 101:365–376, 1987

    Google Scholar 

  50. Takahashi K, Kako KJ: The effect of myocardial ischemia and Nisoldipine pretreatment on the asymmetric distribution of phosphatidylethanolamine in a canine heart sarcolemmal preparation. Biochem Med Met Biol 35:308–321, 1986

    Google Scholar 

  51. Kako KJ: Isolation of sarcolemmal membrane, membrane orientation, and lipid asymmetry. Med Res Rev 7:507–522, 1987

    Google Scholar 

  52. Malaisse WJ: Alloxan toxicity to the pancreatic B-cell. A new hypothesis. Biochem Pharmacol 31:3527–3534, 1982

    Google Scholar 

  53. Black HE, Rosemblum TY. Chemically induced diabetes mellitus in the dog. Am J Pathol. 98:295–310, 1980

    Google Scholar 

  54. McEvoy RC, Hegre OD: Morphometric quantitation of the pancreatic insulin-, glucagon and somatostatin-positive cell populations in normal and alloxan diabetic rats. Diabetes 26:1140–1146, 1977

    Google Scholar 

  55. Steinman RM: Endocytosis and the recycling of plasma membrane. J Cell Biol 96:1–27, 1983

    Google Scholar 

  56. Shamoo AE, Ambudkar IS: Regulation of calcium transport in cardiac cells. Can J Physiol Pharmacol 62:9–22, 1984

    Google Scholar 

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  1. Department of Physiology, Faculty of Health Sciences, University of Ottawa, Smyth Road, KIH 8M5, Ottawa, Ontario, Canada

    Mitsutoshi Kato & K. Joe Kako

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  1. Mitsutoshi Kato
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  2. K. Joe Kako
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Kato, M., Kako, K.J. Na+/Ca2+ exchange of isolated sarcolemmal membrane: effects of insulin, oxidants and insulin deficiency. Mol Cell Biochem 83, 15–25 (1988). https://doi.org/10.1007/BF00223194

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  • DOI: https://doi.org/10.1007/BF00223194

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