title: 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.
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.
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Universidade de São Paulo, FCFRP-USP, Ribeirão Preto, SP, Brazil.
authors.forename: Tassia R
authors.initials: TR
authors.lastname: Costa
authors.forename: Danilo L
authors.initials: DL
authors.lastname: Menaldo
authors.forename: Clayton Z
authors.initials: CZ
authors.lastname: Oliveira
authors.forename: Norival A
authors.initials: NA
authors.lastname: Santos-Filho
authors.forename: Sabrina S
authors.initials: SS
authors.lastname: Teixeira
authors.forename: Auro
authors.initials: A
authors.lastname: Nomizo
authors.forename: André L
authors.initials: AL
authors.lastname: Fuly
authors.forename: Marta C
authors.initials: MC
authors.lastname: Monteiro
authors.forename: Bibiana M
authors.initials: BM
authors.lastname: de Souza
authors.forename: Mário S
authors.initials: MS
authors.lastname: Palma
authors.forename: Rodrigo G
authors.initials: RG
authors.lastname: Stábeli
authors.forename: Suely V
authors.initials: SV
authors.lastname: Sampaio
authors.forename: Andreimar M
authors.initials: AM
authors.lastname: Soares
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D045744:Cell Line, Tumor / Q000187:drug effects*; D003435:Crotalid Venoms / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D051379:Mice; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D054467:Phospholipases A2 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D003435:Crotalid Venoms; D007527:Isoenzymes; D010455:Peptides; D054467:Phospholipases A2