A new L-amino acid oxidase from Bothrops jararacussu snake venom: Isolation, partial characterization, and assessment of pro-apoptotic and antiprotozoal activities

作者：Carone Sante E I; Costa Tassia R; Burin Sandra M; Cintra Adelia C O; Zoccal Karina F; Bianchini Francine J; Tucci Luiz F F; Franco Joao J; Torqueti Maria R; Faccioli Lucia H; de Albuquerque Sergio; de Castro Fabiola A; Sampaio Suely V^*

来源：International Journal of Biological Macromolecules, 2017, 103: 25-35.

DOI：10.1016/j.ijbiomac.2017.05.025

• Snake venom
• Bothrops jararacussu
• L-amino acid oxidase
• Apoptosis
• Antiprotozoal activity

A new L-amino acid oxidase (LAAO) from Bothrops jararacussu venom (BjussuLAAO-II) was isolated by using a three-step chromatographic procedure based on molecular exclusion, hydrophobicity, and affinity. BjussuLAAO-II is an acidic enzyme with pI = 3.9 and molecular mass = 60.36 kDa that represents 0.3% of the venom proteins and exhibits high enzymatic activity (4884.53 U/mg/mim). We determined part of the primary sequence of BjussuLAAO-II by identifying 96 amino acids, from which 34 compose the N-terminal of the enzyme (ADDRNPLEECFRETDYEEFLEIARNGLSDTDNPK). Multiple alignment of the partial BjussuLAAO-II sequence with LAAOs deposited in the NCBI database revealed high similarity (95-97%) with other LAAOs isolated from Bothrops snake venoms. BjussuLAAO-II exerted a strong antiprotozoal effect against Leishmania amazonensis (IC50 = 4.56 mu g/mL) and Trypanosoma cruzi (IC50 = 4.85 mu g/mL). This toxin also induced cytotoxicity (IC50 = 1.80 mu g/mL) and apoptosis in MCF7 cells (a human breast adenocarcinoma cell line) by activating the intrinsic and extrinsic apoptosis pathways, but were not cytotoxic towards MCF10A cells (a non-tumorigenic human breast epithelial cell line). The results reported herein add important knowledge to the field of Toxinology, especially for the development of new therapeutic agents.

• 出版日期2017-10