Journal of Protein Chemistry

, Volume 21, Issue 3, pp 131–136 | Cite as

Isolation and Preliminary Enzymatic Characterization of a Novel PLA2 from Crotalus durissus collilineatus Venom

  • L. A. Ponce-Soto
  • M. H. Toyama
  • S. Hyslop
  • J. C. Novello
  • S. Marangoni


A crotoxin homolog was purified from the Crotalus durissus collilineatus venom using molecular exclusion and reverse-phase HPLC. This crotoxin contained one PLA2 (Cdcolli III F6) and four crotapotin isoforms, whereas crotoxin from Crotalus durissus terrificus venom had three PLA2 isoforms and two crotapotin isoforms. SDS-PAGE showed that the C. d. collilineatus PLA2 and crotapotin had relative molecular mass of 15 and 9 kDa, respectively. Neither the PLA2 (Cdcolli III F6) nor the crotapotins (Cdcolli III F3 and F4) had any neurotoxicity in mouse phrenic nerve-diaphragm preparations when tested alone. However, when PLA2 and crotapotin were coincubated before testing, the neurotoxicity was restored to a level similar to test in the venom in native crotoxin. The two crotapotins (Cdcolli III F3 and F4) differed in their ability to inhibit PLA2 activity, perhaps because of variations in their affinities for this enzyme. Cdcolli III F6 showed allosteric enzymatic behavior, with maximal activity at pH 8.3 and 36°C. Full PLA2 activity required the presence of a low Ca2+ concentration and was inhibited by Cu2+ and Zn2+ and by Cu2+ and Mg2+ in the presence and absence of Ca2+, respectively. These results indicate that crotoxin from C. d. collineatus venom is very similar enzymatically to crotoxin from C. d. terrificus.

Crotalus durissus collilineatus crotapotin crotoxin phospholipase A2 rattlesnake venom 


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  1. Azevedo-Marques, M. M., Cupo, P., Coimbra, T. M., Hering, S. E., Rossi, M. A., and Laure, C. J. (1982). Toxicon 23, 631-636.Google Scholar
  2. Beghini, D. G., Toyama, M. H., Hyslop, S., Sodek, L. C., Novello, J. C., and Marangoni, S. (2000). J. Prot. Chem. 19, 679-664.Google Scholar
  3. Bon, C., Changeux, J. P., Jeng, T. W., and Fraenkel-Conrat, H. (1979). Eur. J. Biochem. 99, 471-481.Google Scholar
  4. Bonfim, V. L., Toyama, M. H., Novello, J. C., Hyslop, S., Oliveira, C. R. B., Rodrigues-Simioni, L., and Marangoni, S. (2001). J. Prot. Chem. 20, 239-245.Google Scholar
  5. Breithaupt, H. (1976). Toxicon 14, 221-233.Google Scholar
  6. Cho, W. and Kezdy, F. J. (1991). Methods Enzymol. 197, 75-79.Google Scholar
  7. Cupo, P., Azevedo-Marques, M. M., and Hering, S. E. (1988). Trans. R. Soc. Trop. Med. Hyg. 82, 924-929.Google Scholar
  8. Faure, G. and Bon, C. (1987). Toxicon 25, 229-234.Google Scholar
  9. Faure, G. and Bon, C. (1988). Biochemistry 27, 730-738.Google Scholar
  10. Faure, G., Choumet, V., Bouchier, C., Camoin, L., Guillaume, J. L., Monegier, B., Vuilhorgne, M., and Bon, C. (1994). Eur. J. Biochem. 223, 161-164.Google Scholar
  11. Gutiérrez, J. M. and Lomonte, B. (1995). Toxicon 33, 1405-1424.Google Scholar
  12. Habermann, E. and Breithaupt, H. (1978). Toxicon 16, 19-30.Google Scholar
  13. Hendon, R. A. and Fraenkel-Conrat, H. (1971). Proc. Natl. Acad. Sci. USA 68, 1560-1563.Google Scholar
  14. Holzer, M. and Mackessy, S. P. (1996). Toxicon 34, 1149-1155.Google Scholar
  15. Landucci, E. C. T., Condino-Neto, A., Perez, A. C., Hyslop, S., Corrado, A. P., Novello, J. C., Marangoni, S., Oliveira, B., Antunes, E., and de Nucci, G. (1994). Toxicon 32, 217-226.Google Scholar
  16. Lennon, B. W. and Kaiser, I. (1990). Comp. Biochem. Physiol. B. 97, 695-699.Google Scholar
  17. Pieterson, W. A., Volwerck, J. J., and de Haas, G. H. (1974). Biochemistry 13, 1439-1445.Google Scholar
  18. Rosenfeld, G. (1971). In Venomous animals and their venoms. Vol. II. (Bücherl, W., and Buckley, E. E., eds), Academic Press, New York, pp. 345-362.Google Scholar
  19. Rubsamen, K., Breithaupt, H., and Habermann, E. (1971). Naunyn-Schmiedeberg's Arch. Pharmacol. 270, 274-288.Google Scholar
  20. Schägger, H. and Von Jagow, G. (1987). Anal. Biochem. 166, 368-379.Google Scholar
  21. Toyama, M. H., Mancuso, L. C., Giglio, J. R., Novello, J. C., Oliveira, B., and Marangoni, S. (1995). Biochem. Mol. Biol. Int. 37, 1047-1055.Google Scholar
  22. Toyama, M. H., Carneiro, E. M., Marangoni, S., Barbosa, R. L., Corso, G., and Boschero, A. C. (2000). Biochim. Biophys. Acta 1474, 56-60.Google Scholar
  23. Tu, A. T., Passey, R. B., and Toom, P. M. (1970). Arch. Biochem. Biophys. 140, 96-106.Google Scholar

Copyright information

© Plenum Publishing Corporation 2002

Authors and Affiliations

  • L. A. Ponce-Soto
    • 1
  • M. H. Toyama
    • 1
    • 2
  • S. Hyslop
    • 3
  • J. C. Novello
    • 1
  • S. Marangoni
    • 1
  1. 1.Universidade Estadual de Campinas (UNICAMP)Campinas, SPBrasil
  2. 2.Departamento de Fisiologia, Instituto de BiologiaUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrasil
  3. 3.Departamento de Farmacologia, Faculdade de Ciências MédicasUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil

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