Myotoxic phospholipases A(2) isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: cytotoxic effect on microorganism and tumor cells

Peptides. 2008 Oct;29(10):1645-56. doi: 10.1016/j.peptides.2008.05.021. Epub 2008 Jun 5.

Abstract

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A(2) (PLA(2)s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing M(r) approximately 14,000 for the monomer and 28,000Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA(2)s from snake venoms, MTX-I belonging to Asp49 PLA(2) class, enzymatically active, and MTX-II to Lys49 PLA(2)s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA(2) and anticoagulant activities, corroborating the importance of residue His48 and Ca(2+) ions for the enzymatic catalysis. Both PLA(2)s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA(2) proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bothrops / metabolism*
  • Cell Line, Tumor / drug effects*
  • Crotalid Venoms / enzymology*
  • Humans
  • Isoenzymes / genetics
  • Isoenzymes / isolation & purification
  • Isoenzymes / metabolism*
  • Male
  • Mice
  • Microbial Sensitivity Tests
  • Molecular Sequence Data
  • Peptide Mapping
  • Peptides / genetics
  • Peptides / isolation & purification
  • Peptides / pharmacology*
  • Phospholipases A2 / genetics
  • Phospholipases A2 / isolation & purification
  • Phospholipases A2 / metabolism*
  • Sequence Alignment
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Crotalid Venoms
  • Isoenzymes
  • Peptides
  • Phospholipases A2