Artículos de revistas
Evidence of caspase-mediated apoptosis induced by L-amino acid oxidase isolated from Bothrops atrox snake venom
Fecha
2008Registro en:
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY, v.151, n.4, p.542-550, 2008
1095-6433
10.1016/j.cbpa.2008.07.007
Autor
ALVES, Raquel Melo
ANTONUCCI, Gilmara Ausech
PAIVA, Helder Henrique
CINTRA, Adelia Cristina Oliveira
FRANCO, Joao Jose
MENDONCA-FRANQUEIRO, Elaine Paula
DORTA, Daniel Junqueira
GIGLIO, Jose Roberto
ROSA, Jose Cesar
FULY, Andre Lopes
DIAS-BARUFFI, Marcelo
SOARES, Andreirnar Martins
SAMPAIO, Suely Vilela
Institución
Resumen
The aim of this work was to investigate the involvement of caspases in apoptosis induced by L-amino acid oxidase isolated from Bothrops atrox snake venom. The isolation of LAAO involved three chromatographic steps: molecular exclusion on a G-75 column; ion exchange column by HPLC and affinity chromatography on a Lentil Lectin column. SDS-PAGE was used to confirm the expected high purity level of BatroxLAA0. It is a glycoprotein with 12% sugar and an acidic character, as confirmed by its amino acid composition, rich in ""Asp and Glu"" residues. It displays high specificity toward hydrophobic L-amino acids. The N-terminal amino acid sequence and internal peptide sequences showed close structural homology to other snake venom LAAOs. This enzyme induces in vitro platelet aggregation, which may be due to H(2)O(2) production by LAAOs, since the addition of catalase completely inhibited the aggregation effect. It also showed cytotoxicity towards several cancer cell lines: HL60, Jurkat, B16F10 and PC12. The cytotoxicity activity was abolished by catalase. A fluorescence microscopy evaluation revealed a significant increase in the apoptotic index of these cells after BatroxLAAO treatment. This observation was confirmed by phosphatidyl serine exposure and activation of caspases. BatroxLAAO is a protein with various biological functions that can be involved in envenomation. Further investigations of its function will contribute to toxicology advances. Published by Elsevier Inc.