Skip to main content
SpringerLink
Log in

Effects of free radicals on cytosolic creatine kinase activities and protection by antioxidant enzymes and sulfhydryl compounds

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The main purpose of this study was to investigate the effect of free radicals and experimental diabetes on cytosolic creatine kinase activity in rat heart, muscle and brain. Hydrogen peroxide decreased creatine kinase activity in a dose dependent manner which was reversed by catalase. Xanthine/xanthine oxidase, which produces superoxide anion, lowered the creatine kinase activity in the same manner whose effect was protected by superoxide dismutase. N-acetylcysteine and dithiothreitol also significantly ameliorated the effect of Xanthine/xanthine oxidase and hydrogen peroxide. Experimental diabetes of twenty-one days (induced by alloxan), also caused a similar decrease in the activity of creatine kinase. This led us to the conclusion that the decrease in creatine kinase activity during diabetes could be due to the production of reactive oxygen species. The free radical effect could be on the sulfhydryl groups of the enzyme at the active sites, since addition of sulfhydryl groups like N-acetylcysteine and dithiothreitol showed a significant reversal effect.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bessman SP, Geiger PJ: The phosphoryl creatine shuttle. Science 211: 448–452, 1981

    Google Scholar 

  2. Wallimann T, Schlosser T, Eppenberger HM: Function of M-line bound CK as intramyofibrillar ATP regenerator at the receiving end of the phosphoryl creatine shuttle in muscle. J Biol Chem 259: 5224–5238, 1984

    Google Scholar 

  3. Yagi K, Mase R: Coupled reaction of creatine kinase and myosin ATPase. J Biol Chem 237: 397–403, 1962

    Google Scholar 

  4. Savabi F, Gieger PJ, Bessman SP: Kinetic properties and functional role of creatine phosphokinase in glycerinated muscle fibers, further evidence for compartmentation. Biochem Biophys Res Commun 114: 785–790, 1993

    Google Scholar 

  5. Wallimann T, Wyss M, Bridzka D, Nicolay K, Eppenberger HM: Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: The ‘Phosphocreatine circuit’ for cellular energy homeostasis. Biochem J 81: 21–40, 1992

    Google Scholar 

  6. Kaldis P, Wallimann T: Functional differences between dimeric and octameric mitochondrial creatine kinase. Biochem J 388: 623–627, 1995

    Google Scholar 

  7. Przyklenk K, Whittaker P, Kloner RA: In vivo infusion of oxygen free radicals. Am Heart J 119: 807–815, 1990

    Google Scholar 

  8. Wei Z, Benjamin HSL: Garlic inhibits free radical generation and augments antioxidant enzyme activity in vascular endothelial cells. Nutr Res 18: 61–70, 1998

    Google Scholar 

  9. Oberley LW: Free radicals and diabetes. Free Radic Biol Med 5: 113–124, 1988

    Google Scholar 

  10. Crawford DW, Blankenhorn DH: Arterial wall oxygenation, oxyradicals and atherosclerosis. 89: 97–108, 1991

    Google Scholar 

  11. Kenyon GL: Creatine kinase shapes up. Nature 381: 281–282, 1996

    Google Scholar 

  12. Haghighi AZ, Maples KR: On the mechanism of the inhibition of glutamine synthetase and creatine phosphokinase by methionine sulfoxide. J Neurosci Res 43: 107–111, 1996

    Google Scholar 

  13. Konorev EA, Hogg N, Kalyanaraman B: Rapid and irreversible inhibition of creatine kinase by peroxynitrite. FEBS Lett 427: 171–174, 1998

    Google Scholar 

  14. Halliwell B, Gutteridge JMC: Lipid peroxidation, oxygen radicals, cell damage and antioxidant therapy. Lancet 1: 1396–1397, 1984

    Google Scholar 

  15. Heikila RE, Winston B, Cohen G, Barden H: Alloxan induced diabetes, evidence for hydroxyl radical as a toxic intermediate. Biochem Pharmacol 25: 1085–1092, 1976

    Google Scholar 

  16. Sochor M, Baquer NZ, McLean P: Glucose over and under utilization in diabetes: Comparative studies on the change in activities of enzymes of glucose metabolism in rat kidney and liver. Mol Physiol 7: 51–68.

  17. Forster G, Bernt E, Bergmeyer HU: In HU Bergmeyer (ed.) Methods in Enzymology. vol. II. Academic Press, New York, 1974, pp 784–793

    Google Scholar 

  18. Scherer NM, Deamer DW: Oxidative stress impairs the function of sarcoplasmic reticulum by oxidation of sulfhydryl groups in the Ca2+ ATPase. Arch Biochem 246: 589–610, 1986

    Google Scholar 

  19. Lowry H, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  20. Genet S, Kale RK, Baquer NZ: Effect of vanadate, insulin and fenugreek (Trigonella foenum graecum) on creatine kinase levels in tissues of diabetic rat. Ind J Exp Biol 37: 200–202, 1999

    Google Scholar 

  21. Zhang Y, Marcillar O, Giulivic C, Ernster L, Davies KJA: The oxidative inactivation of mitochondrial transport chain components and ATPase. J Biol Chem 265: 16330–16336, 1990

    Google Scholar 

  22. Liu P, Hock CE, Nagele R, Wong PY: Formation of nitricoxide, superoxide and peroxynitrite in myocardial ischemia-reperfusion injury in rats. Am J Physiol 272(5pt2): H2327–H2336, 1997

    Google Scholar 

  23. Liu ZJ, Zhou J: Spin-labeling probe on conformational change at the active site of creatine kinase during denaturation by guanidine hydrochloride. Biochem Biophys Acta 1253: 63–68, 1995

    Google Scholar 

  24. Yatin SM, Aksenov M, Butterfield DA: The antioxidant vitamin E modulates amyloid beta-peptide induced creatine kinase activity inhibition and increased protein oxidation: Implications for the free radical hypothesis of Alzheimer's disease. Neurochem Res 24: 427–435, 1999

    Google Scholar 

  25. Ventura-Clapier R, Veksler V, Hoerter JA: Myofabrillar creatine kinase and cardiac contraction. Mol Cell Biochem 133/134: 1125–1144, 1994

    Google Scholar 

  26. Tian R, Ingwall JS: Energetic basis for reduced contractile reserve in isolated rat hearts. Am J Physiol 270: H1207–H1216, 1996

    Google Scholar 

  27. Veksler V, Ventura-Clapier R: In situ study of myofibrils, mitochondria and bound creatine kinase in experimental cardiomyopathies. Mol Cell Biochem 133/134: 287–298, 1994

    Google Scholar 

  28. Mathews RT, Yang L, Jenkins GG, Ferrants RJ, Rosen BR, Kaddurah-daouk R, Beal MF: Sustrates cause myocardial systolic but not diastolic disfunction. J Neurosci 18: 156–163, 1998

    Google Scholar 

  29. Mekhfi H, Veksler V, Mateo P, Rochette L, Ventura-Clapier R: Creatine kinase is the main target of reactive oxygen species in cardiac myofibrils. Circ Res 78: 1016–1027, 1996

    Google Scholar 

  30. Baynes, JW: Role of oxidative stress in development of complications in diabetes. Diabetes 40: 405–412, 1991

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Hormone and Drug Research Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Dehli, India

    Solomon Genet, Raosaheb Kale & Najma Baquer

Authors
  1. Solomon Genet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Raosaheb Kale
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Najma Baquer
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Genet, S., Kale, R. & Baquer, N. Effects of free radicals on cytosolic creatine kinase activities and protection by antioxidant enzymes and sulfhydryl compounds. Mol Cell Biochem 210, 23–28 (2000). https://doi.org/10.1023/A:1007071617480

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007071617480

Search

Navigation