Structural and functional properties of BaTX, a new Lys49 phospholipase A2 homologue isolated from the venom of the snake Bothrops alternatus

Luis Alberto Ponce-Soto, Bruno Lomonte, José María Gutiérrez, Lea Rodrigues-Simioni, José Camillo Novello, Sergio Marangoni

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

BaTX PLA2, a K49 phospholipase A2 homologue was purified from Bothrops alternatus venom after two chromatographic steps, molecular exclusion on Superdex 75 and reverse phase HPLC on μ-Bondapack C-18. A molecular mass of 13898.71 Da was determined by MALDI-TOF mass spectrometry. The amino acid composition showed that BaTX has a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic PLA2. The complete amino acid sequence of BaTX PLA2 contains 121 residues, resulting in a calculated pI value of 8.63. This sequence shows high identity values when compared to other K49 PLA2s isolated from the venoms of viperid snakes. Lower identity is observed in comparison to D49 PLA2s. The sequence was SLFELGKMIL QETGKNPAKS YGAYYCYCGW GGQGQPKDAT DRCCYVHKCC YKKLTGCNPK KDRYSYSWKD KTIVCGENNS CLKELCECDK AVAICLRENL NTYNKKYRYY LKPLCKKADA C. In mice, BaTX induced myonecrosis and edema, upon intramuscular or subcutaneous injections, respectively. The LD50 of BaTX was 7 μg/g body weight, by intravenous route. In vitro, the toxin caused a potent blockade of neuromuscular transmission in young chicken biventer cervicis preparations. The blockage 50% was achieved at a concentration of 0.03 μM: 40 ± 0.4 min and 0.07 μM: 35 ± 0.3 min. Moreover, this protein induced a rapid cytolytic effect upon mouse skeletal muscle myoblasts in culture. Thus, the combined structural and functional information obtained identify BaTX as a new member of the K49 PLA2 family, which presents the typical bioactivities described for such proteins.

Original languageEnglish
Pages (from-to)585-593
Number of pages9
JournalBiochimica et Biophysica Acta - General Subjects
Volume1770
Issue number4
DOIs
StatePublished - Apr 2007
Externally publishedYes

Keywords

  • Bothrops alternatus
  • Lys-49
  • Myotoxin
  • Neurotoxin
  • Phospholipase A
  • Snake venom

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