Journal of Protein Chemistry

, Volume 19, Issue 8, pp 679–684

Enzymatic Characterization of a Novel Phospholipase A2 from Crotalus durissus cascavella Rattlesnake (Maracambóia) Venom

  • D. G. Beghini
  • M. H. Toyama
  • S. Hyslop
  • L. C. Sodek
  • Novello
  • S. Marangoni
Article

DOI: 10.1023/A:1007152303179

Cite this article as:
Beghini, D.G., Toyama, M.H., Hyslop, S. et al. J Protein Chem (2000) 19: 679. doi:10.1023/A:1007152303179

Abstract

The PLA2 and crotapotin subunits of crotoxin from Crotalus durissus cascavella venom were purified by a combination of high-performance liquid chromatography (HPLC) molecular exclusion (Protein Pack 300SW column) and reverse-phase HPLC (RP-HPLC). Tricine SDS-PAGE showed that the PLA2 and crotapotins migrated as single bands with estimated molecular masses of 15 and 9 kDa, respectively. The amino acid composition of the PLA2 showed the presence of 14 half-cysteines and a high content of basic residues (Lys, Arg, His), whereas the crotapotins were rich in hydrophobic, negatively charged residues and half-cysteines. The PLA2 showed allosteric behavior, with maximal activity at pH 8.3 and 35–40°C. C. d. cascavella PLA2 required Ca2+ for activity but was inhibited by Cu2+ and Zn2+ and by Cu2+ and Mg2+ in the presence and absence of Ca2+, respectively. Crotapotin (F3) and heparin inhibited the catalytic activity of the PLA2 by acting as allosteric inhibitors.

Crotalus durissus cascavella crotoxin crotapotin PLA2 rattlesnake venom 

Copyright information

© Plenum Publishing Corporation 2000

Authors and Affiliations

  • D. G. Beghini
    • 1
  • M. H. Toyama
    • 1
    • 2
  • S. Hyslop
    • 3
  • L. C. Sodek
    • 2
  • Novello
    • 1
  • S. Marangoni
    • 4
  1. 1.Departamento de Bioquímica, Instituto de BiologiaUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil
  2. 2.Departamento de Fisiologia, Instituto de Biologia (IB), (Univesidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil
  3. 3.Departamento de Farmacologia, Faculdade de Ciências MédicasUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil
  4. 4.Departamento de Bioquímica, Instituto de BiologiaUniversidade Estadual de Campinas (UNICAMP)Campinas, SPBrazil