The Protein Journal

, Volume 24, Issue 2, pp 103–112

Biological and Structural Characterization of a New PLA2 from the Crotalus durissus collilineatus Venom

  • M. H. Toyama
  • D. O. Toyama
  • Paulo P. Joazeiro
  • E. M. Carneiro
  • L. O. S. Beriam
  • L. S. Marangoni
  • A. C. Boschero
Article

DOI: 10.1007/s10930-004-1517-5

Cite this article as:
Toyama, M.H., Toyama, D.O., Joazeiro, P.P. et al. Protein J (2005) 24: 103. doi:10.1007/s10930-004-1517-5

Abstract

In the present article we report on the biological characterization and amino acid sequence of a new basic Phospholipases A2 (PLA2) isolated from the Crotalus durissus collilineatus venom (Cdcolli F6), which showed the presence of 122 amino acid residues with a pI value of 8.3, molecular mass of 14 kDa and revealed an amino acid sequence identity of 80 with crotalic PLA2s such as Mojave B, Cdt F15, and CROATOX. This homology, however, dropped to 50 if compared to other sources of PLA2s such as from the Bothrops snake venom. Also, this PLA2 induced myonecrosis, although this effect was lower than that of BthTx-I or whole crotoxin and it was able to induce a strong blockage effect on the chick biventer neuromuscular preparation, independently of the presence of the acid subunid (crotapotin). The neurotoxic effect was strongly reduced by pre-incubation with heparin or with anhydrous acetic acid and q-BPB showed a similar reduction. The q-BPB did not reduce significantly the myotoxic activity induced by the PLA2, but the anhydrous acetic acid treatment and the pre-incu-bation of PLA2 with heparin reduced significantly its effects. This protein showed a strong antimicrobial activity against Xanthomonas axonopodis passiflorae (Gram-negative), which was drastically reduced by incubation of this PLA2 with q-BPB, but this effect was marginally reduced after treatment with anhydrous acetic acid. Our findings here allow to speculate that basic amino acid residues on the C-terminal and molecular regions near catalytic site regions such as Calcium binding loop or b-wing region may be involved in the binding of this PLA2 to the molecular receptor to induce the neurotoxic effect. The bactericidal effect, however, was completely dependent on the enzymatic activity of this protein.

Keywords

Crotalus durisssus collilineatus Crotoxin myonecrosis neurotoxicity PLA2 rattlesnake 

Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • M. H. Toyama
    • 1
  • D. O. Toyama
    • 2
  • Paulo P. Joazeiro
    • 3
  • E. M. Carneiro
    • 4
  • L. O. S. Beriam
    • 5
  • L. S. Marangoni
    • 1
  • A. C. Boschero
    • 4
  1. 1.Campus do Litoral Paulista, Unidade de São VicenteUNESPBrazil
  2. 2.Departamento de BioquímicaInstituto de Biologia, UNICAMPBrazil
  3. 3.Departamento de Histologia e EmbriologiaInstituto de Biologia, UNICAMPBrazil
  4. 4.Departamento de Fisiologia e BiofísicaInstituto de Biologia, UNICAMPBrazil
  5. 5.Departamento de MicrobiologiaInstituto Biológico, CampinasSao PauloBrazil