Journal of Pharmacopuncture (대한약침학회지)

KOREAN PHARMACOPUNCTURE INSTITUTE (대한약침학회)
권호 표지
DOI QR코드

DOI QR Code

Purification and Characterization of a Fibrinolytic Enzyme from Snake Venom of Macrovipera lebetina turanica

  • Kwon, Ki-Rok (Graduate School of Oriental Medicine, Sangji University) ;
  • Park, Do-Il (Graduate School of Oriental Medicine, Sangji University) ;
  • Lee, Seung-Bae (Division of Animal Resources and Life Science, Sangji University) ;
  • Choi, Suk-Ho (Division of Animal Resources and Life Science, Sangji University)
  • Received : 2010.12.23
  • Accepted : 2011.04.19
  • Published : 2011.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Objectives: Fibrinolytic enzyme preparations were isolated from the snake venom of Macrovipera lebetica turanica in this study. Methods: The purity of the preparations was determined using SDS-PAGE and the enzymic characteristics of the purified fibrinolytic enzyme were determined. Results: 1. All of the two preparations with fibrinolytic activity obtained from the snake venom of M. l. turanicat contained the major polypeptide with the molecular weight of 27,500. One of the preparation showed purified fibrinolytic enzyme. 2. The purified fibrinolytic enzyme hydrolyzed ${\alpha}$-chain of fibrinogen faster than ${\beta}$-chain but not ${\gamma}$-chain. 3. The fibrinolytic activity was inhibited completely by EDTA, EGTA, 1,10-phenanthroline, and dithiothreitol. 4. The fibrinolytic activity was inhibited completely by calcium chloride, iron(III) chloride, mercuric chloride, and cobalt (II) chloride. 5. The fibrinolysis zone formed after addition of zinc sulfate was smaller but clearer than the control. Conclusions: These results suggested that the fibrinolytic enzyme purifed from the snake venom of M. l turanica was a metalloprotease containing dithiol group.

Keywords

References

  1. Markland, F. S. : Snake venoms and the hemostatic system. Toxicon 36:1749-1800, 1998. https://doi.org/10.1016/S0041-0101(98)00126-3
  2. Russel, F. E. : Toxic effects of animal toxins. In Klassen, C.D., Amdur, M.O., and Doull, J. (Ed.) Casarett and Doull's toxicology-the basic science of poisons, 3rd ed. MacMillan, New York, pp. 706-756, 1986.
  3. Ramos, O. H. P. and Selistre-de-Araujo, H. S. : Snake venome metalloproteases-structure and function of catalytic and disintegrin domains. Comp. Biochem. Physiol. Part C. 142:328-346, 2006.
  4. Marsh, N. and Willams, V. : Practical applications of snake venom toxins in haemostasis. Toxicon 45:1171-1181, 2005. https://doi.org/10.1016/j.toxicon.2005.02.016
  5. Kamiguti, A. S., Hay, C. R. M., Theakston, R. D. G., and Zuzel, M. : Insights into the mechanism of haemorrhage caused by snake venom metalloproteinases. Toxicon 34:627-642, 1996. https://doi.org/10.1016/0041-0101(96)00017-7
  6. Faiz, M. A., Falkous, G., Harris, J. B., and Mantle, D. : Comparison of protease and related enzyme activities in snake venoms. Comp. Biochem. Physiol. B. 113:199-204, 1996. https://doi.org/10.1016/0305-0491(95)02050-0
  7. Marsh, N. A. : Snake venoms affecting the haemostatic mechanism - a consideration of their mechanisms, practical applications and biological significance. Blood Coagul. Fibrinolysis. 5:399-410, 1994.
  8. Samsa, G. P., Matchar, D. B., Williams, G. R., Levy, D. E. : Cost-effectiveness of ancrod treatment of acute ischaemic stroke: results from the Stroke Treatment with Ancrod Trial (STAT). J. Eval. Clin. Pract. 8:61-70, 2002. https://doi.org/10.1046/j.1365-2753.2002.00315.x
  9. Markland, F. S., Reddy, K. N. N., and Guan, L. F. : Purification and characterization of a directacting fibrinolytic enzyme from Southern copperhead venom. In: Pirkle, H., Markland, F. S. (Eds.), Hemostasis and Animal Venoms. Marcel Dekker, New York, pp. 173-189, 1988.
  10. Swenson, S. and Markland Jr. F. S. : Snake venom fibrin(ogen)olytic enzymes. Toxicon 45:1021-1039, 2005. https://doi.org/10.1016/j.toxicon.2005.02.027
  11. Toombs, C. F. : Alfimplrase: pharmacology of a novel fibrinolytic metalloproteinase for thrombolysis. Haemostasis 31:141-147, 2001.
  12. Baker, B. J., Tu, A. T. : Atroxase: a fibrinolytic enzyme isolated from the venom of western diamondback rattlesnake: isolation, characterization and cloning. Adv. Exp. Med. Biol. 391:203-211, 1996. https://doi.org/10.1007/978-1-4613-0361-9_13
  13. Gasmi, A., Chabchoub, A., Guermazi, S., Karoui, H., Elayeb, M., and Dellagi, K. : Further characterization and thrombolytic activity in a rat model of a fibrinogenase from Vipera lebetina venom. Thromb. Haemostas. 86:233-242, 1997.
  14. McDiarmid, R. W., Campbell, J. A., and Toure, T. : Snake species of the world: Taxonomic and geographic reference, vol. 1. Herpetologists League pp. 511, 1999.
  15. Integrated Taxonomic Information System : Macrovipera labetina.(http://www.itis.gov/servlet/SingleRpt/SingleRpt?search_topic=TSN&search_value=634977), 2009.
  16. Siigur, J., Aaspollu, A., Tonismagi, K., Trummal, K., Samel, M., Vija, H., Subbi, J., Siigur, E., : Proteases from Vipera lebetina venom affecting coagulation and fibrinolysis. Haemostasis 31:123-132, 2001.
  17. Siigur, E. and Siigur, J. Purification and characterization of lebetase, a fibrinolytic enzyme from Vipera lebetina (snake) venom. Biochim. Biophys. Acta. 1074:223-229, 1991. https://doi.org/10.1016/0304-4165(91)90156-B
  18. Siigur, J., Samel, M., Tonismagi, K., Subbi, J., and Siigur, E. : Biochemical characterization of lebetase, a direct-acting fibrinolytic enzyme from Vipera lebetina snake venom. Thromb. Res. 90:39-49, 1998. https://doi.org/10.1016/S0049-3848(98)00009-7
  19. Siigur, E., Aaspollu, A., Tu, A. T., and Siigur, J. : cDNA cloning and deduced amino acid sequence of fibrinolytic enzyme (lebetase) from Vipera lebetina snake venom. Biochem. Biophy. Res. Comm. 224:297-2236, 1996. https://doi.org/10.1006/bbrc.1996.1024
  20. Astrup, T., and Mullertz, S. : The fibrin plate method for estimating of fibrinolytic activity. Archs. Biochem. Biophys. 40:346-351, 1952. https://doi.org/10.1016/0003-9861(52)90121-5
  21. Laemmli, U. K. : Cleavage of structural proteins during the assembly of the head of bacteriophage T-4. Nature (London) 227: 680-685, 1970. https://doi.org/10.1038/227680a0