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.
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.
Costa
Tassia R
TR
Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Universidade de São Paulo, FCFRP-USP, Ribeirão Preto, SP, Brazil.
Menaldo
Danilo L
DL
Oliveira
Clayton Z
CZ
Santos-Filho
Norival A
NA
Teixeira
Sabrina S
SS
Nomizo
Auro
A
Fuly
André L
AL
Monteiro
Marta C
MC
de Souza
Bibiana M
BM
Palma
Mário S
MS
Stábeli
Rodrigo G
RG
Sampaio
Suely V
SV
Soares
Andreimar M
AM
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
D003435:Crotalid Venoms; D007527:Isoenzymes; D010455:Peptides; D054467:Phospholipases A2