Biochemical and functional characterization of an L-amino acid oxidase isolated from Bothrops pirajai snake venom.
Izidoro LF,
Ribeiro MC,
Souza GR,
Sant'Ana CD,
Hamaguchi A,
Homsi-Brandeburgo MI,
Goulart LR,
Beleboni RO,
Nomizo A,
Sampaio SV,
Soares AM,
Rodrigues VM.
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, UFU, 38400-902 Uberlândia-MG, Brazil.
In this work we describe the isolation of a new l-amino acid oxidase (LAAO) referred to as BpirLAAO-I from Bothrops pirajai snake venom, which was highly purified using a combination of molecular exclusion, affinity, and hydrophobic chromatography steps. BpirLAAO-I homodimeric acid glycoprotein (approximate Mr and pI of 130,000 and 4.9, respectively) displays high specificity toward hydrophobic/aromatic amino acids, while deglycosylation does not alter its enzymatic activity. The N-terminal LAAO sequence of its first 49 amino acids presented a high similarity between a amino acid sequence with other LAAOs from: Bothrops spp., Crotalus spp., Calloselasma rhodostoma, Agkistrodon spp., Trimeresurus spp., Pseudechis australis, Oxyuranus scutellatus, and Notechis scutatus. BpirLAAO-I induces time-dependent platelet aggregation, mouse paw edema, cytotoxic activity against Escherichia coli, Pseudomonas aeruginosa, Leishmania sp., and tumor cells, and also a typical fago (M13mp18) DNA fragmentation. Platelet aggregation, leishmanicidal and antitumoral activities were reduced by catalase. Thus, BpirLAAO-I is a multifunctional protein with promising biotechnological and medical applications.
PMID: 16809041 [PubMed - indexed for MEDLINE]