Structural and functional properties of Bp-LAAO, a new l-amino acid oxidase isolated from Bothrops pauloensis snake venom

• Published in: Biochimie 2009-04, Vol.91 (4), p.490-501
• Main Authors: Rodrigues, Renata S., da Silva, Juliana F., Boldrini França, Johara, Fonseca, Fernando P.P., Otaviano, Antônio R., Henrique Silva, Flávio, Hamaguchi, Amélia, Magro, Angelo J., Braz, Antônio Sérgio K., dos Santos, Juliana I., Homsi-Brandeburgo, Maria Inês, Fontes, Marcos R.M., Fuly, André L., Soares, Andreimar M., Rodrigues, Veridiana M.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Bothrops ( neuwiedi) pauloensis; Bothrops - metabolism; Breast Neoplasms - metabolism; cDNA library; Cell Line, Tumor; Crotalid Venoms - enzymology; Escherichia coli - drug effects; Humans; l-Amino acid oxidase; L-Amino Acid Oxidase - chemistry; L-Amino Acid Oxidase - metabolism; L-Amino Acid Oxidase - pharmacology; Leishmania - drug effects; Leukemia, T-Cell - metabolism; Molecular dynamics simulation; Molecular Sequence Data; Molecular Structure; Phylogenetic analysis; Phylogeny; Platelet Aggregation - drug effects; Sequence Alignment; Staphylococcus aureus - drug effects; Substrate Specificity - physiology
• ISSN: 0300-9084; 1638-6183
• DOI: 10.1016/j.biochi.2008.12.004

An l-amino acid oxidase (Bp-LAAO) from Bothrops pauloensis snake venom was highly purified using sequential chromatography steps on CM-Sepharose, Phenyl-Sepharose CL-4B, Benzamidine Sepharose and C18 reverse-phase HPLC. Purified Bp-LAAO showed to be a homodimeric acidic glycoprotein with molecular weight around 65 kDa under reducing conditions in SDS-PAGE. The best substrates for Bp-LAAO were l-Met, l-Leu, l-Phe and l-Ile and the enzyme showed a strong reduction of its catalytic activity upon l-Met and l-Phe substrates at extreme temperatures. Bp-LAAO showed leishmanicidal, antitumoral and bactericidal activities dose dependently. Bp-LAAO induced platelet aggregation in platelet-rich plasma and this activity was inhibited by catalase. Bp-LAAO-cDNA of 1548 bp codified a mature protein with 516 amino acid residues corresponding to a theoretical isoelectric point and molecular weight of 6.3 and 58 kDa, respectively. Additionally, structural and phylogenetic studies identified residues under positive selection and their probable location in Bp-LAAO and other snake venom LAAOs (svLAAOs). Structural and functional investigations of these enzymes can contribute to the advancement of toxinology and to the elaboration of novel therapeutic agents.