Laboratory of Biochemistry and Animal Toxins, Institute of Biotechnology, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil.
Laboratory of Osteoimmunology and Tumor Immunology, Institute of Biomedical Sciences, Federal University of Uberlândia - UFU, Uberlândia, MG, Brazil.
Int J Biol Macromol. 2021 Jan 15;167:267-278. doi: 10.1016/j.ijbiomac.2020.11.146. Epub 2020 Nov 24.
This study aims to examine whether two L-amino acid oxidases isolated from Bothrops snake venom (SV-LAAOs) were cytotoxic to Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis, two causative agents of leishmaniasis, which is an endemic disease in tropical and subtropical countries. The SV-LAAOs BjussuLAAO-II and BmooLAAO-II were isolated from Bothrops jararacussu and Bothrops moojeni venom, respectively, through a three-step chromatography process that used molecular exclusion, hydrophobic interaction, and affinity columns. BmooLAAO-II is a new SV-LAAO isoform that we isolated in this study. The purified BjussuLAAO-II and BmooLAAO-II had high L-amino acid oxidase-specific activity: 3481.17 and 4924.77 U/mg/min, respectively. Both SV-LAAOs were strongly cytotoxic to the two Leishmania species, even at low concentrations. At the same concentration, BjussuLAAO-II and BmooLAAO-II exerted different cytotoxic effects on the parasites. We reported for the first time that the SV-LAAOs suppressed cell proliferation and altered the mitochondrial membrane potential of the two Leishmania species. Surprisingly, BjussuLAAO-II increased the intracellular reactive oxygen species production only in L. (L.) amazonensis, while BmooLAAO-II increased the intracellular reactive oxygen species production only in L. (V.) braziliensis, indicating that these SV-LAAOs had a certain specificity of action.
本研究旨在探讨两种从巴西矛头蝮蛇（Bothrops）蛇毒中分离的 L-氨基酸氧化酶（SV-LAAOs）是否对导致利什曼病（leishmaniasis）的两种病原体——引起皮肤利什曼病的美洲利什曼原虫（Leishmania）（Leishmania） amazonensis 和引起黏膜皮肤利什曼病的巴西利什曼原虫（Viannia）（Viannia） braziliensis 具有细胞毒性。SV-LAAOs BjussuLAAO-II 和 BmooLAAO-II 分别从巴西矛头蝮蛇（Bothrops jararacussu）和波氏矛头蝮蛇（Bothrops moojeni）毒液中通过三步色谱法分离，该方法使用分子排阻、疏水相互作用和亲和柱。BmooLAAO-II 是本研究中分离出的一种新的 SV-LAAO 同工酶。纯化的 BjussuLAAO-II 和 BmooLAAO-II 具有高的 L-氨基酸氧化酶特异性活性：分别为 3481.17 和 4924.77 U/mg/min。两种 SV-LAAOs 即使在低浓度下对两种利什曼原虫也具有强烈的细胞毒性。在相同浓度下，BjussuLAAO-II 和 BmooLAAO-II 对寄生虫的细胞毒性作用不同。我们首次报道 SV-LAAOs 抑制了两种利什曼原虫的细胞增殖并改变了线粒体膜电位。令人惊讶的是，BjussuLAAO-II 仅在 L.（L.） amazonensis 中增加了细胞内活性氧物质的产生，而 BmooLAAO-II 仅在 L.（V.） braziliensis 中增加了细胞内活性氧物质的产生，表明这些 SV-LAAOs 具有一定的作用特异性。
