Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Laboratório de Biotecnologia de Proteínas e Compostos Bioativos da Amazônia Ocidental, LaBioProt, Fundação Oswaldo Cruz, FIOCRUZ, Unidade Rondônia, Porto Velho, RO, Brazil; Instituto Federal de Educação, Ciência e Tecnologia de Rondônia, IFRO, Campus Vilhena, Vilhena, RO, Brazil.
Centro de Estudos de Biomoléculas Aplicadas à Saúde, CEBio, Laboratório de Biotecnologia de Proteínas e Compostos Bioativos da Amazônia Ocidental, LaBioProt, Fundação Oswaldo Cruz, FIOCRUZ, Unidade Rondônia, Porto Velho, RO, Brazil; Centro Para el Desarrollo de la Investigación Científica (CEDIC), Asunción, Paraguay.
Chem Biol Interact. 2021 Sep 1;346:109581. doi: 10.1016/j.cbi.2021.109581. Epub 2021 Jul 21.
Bothrops asper is one of the most important snake species in Central America, mainly because of its medical importance in countries like Ecuador, Panama and Costa Rica, where this species causes a high number of snakebite accidents. Several basic phospholipases A (PLAs) have been previously characterized from B. asper venom, but few studies have been carried out with its acidic isoforms. In addition, since snake venom is a rich source of bioactive substances, it is necessary to investigate the biotechnological potential of its components. In this context, this study aimed to carry out the biochemical characterization of PLA isoforms isolated from B. asper venom and to evaluate the antiparasitic potential of these toxins. The venom and key fractions were subjected to different chromatographic steps, obtaining nine PLAs, four acidic ones (BaspAc-I, BaspAc-II, BaspAc-III and BaspAc-IV) and five basic ones (BaspB-I, BaspB-II, BaspB-III, BaspB-IV and BaspB-V). The isoelectric points of the acidic PLAs were also determined, which presented values ranging between 4.5 and 5. The findings indicated the isolation of five unpublished isoforms, four Asp49-PLA, corresponding to the group of acidic isoforms, and one Lys49-PLA-like. Acidic PLAs catalyzed the degradation of all substrates evaluated; however, for the basic PLAs, there was a preference for phosphatidylglycerol and phosphatidic acid. The antiparasitic potential of the toxins was evaluated, and the acidic PLAs demonstrated action against the epimastigote forms of T. cruzi and promastigote forms of L. infantum, while the basic PLAs BaspB-II and BaspB-IV showed activity against P. falciparum. The results indicated an increase of up to 10 times in antiplasmodial activity, when the Asp49-PLA and Lys49-PLA were associated with one another, denoting synergistic action between these PLA isoforms. These findings correspond to the first report of synergistic antiplasmodial action for svPLAs, demonstrating that these molecules may be important targets in the search for new antiparasitic agents.
矛头蝮是中美洲最重要的蛇类之一，主要是因为在厄瓜多尔、巴拿马和哥斯达黎加等国家，这种蛇类导致了大量的蛇咬伤事故。已经从矛头蝮毒液中鉴定出几种基础磷脂酶 A（PLA），但对其酸性同工酶的研究较少。此外，由于蛇毒是生物活性物质的丰富来源，因此有必要研究其成分的生物技术潜力。在这种情况下，本研究旨在对从矛头蝮毒液中分离的 PLA 同工酶进行生化特性分析，并评估这些毒素的抗寄生虫潜力。毒液和关键馏分经过不同的色谱步骤，得到 9 种 PLA，其中 4 种为酸性（BaspAc-I、BaspAc-II、BaspAc-III 和 BaspAc-IV），5 种为碱性（BaspB-I、BaspB-II、BaspB-III、BaspB-IV 和 BaspB-V）。还测定了酸性 PLA 的等电点，范围在 4.5 到 5 之间。结果表明，分离出了 5 种未发表的同工酶，其中 4 种为 Asp49-PLA，属于酸性同工酶组，还有 1 种 Lys49-PLA 样同工酶。酸性 PLA 催化所有评估底物的降解；然而，对于碱性 PLA，优先作用于磷脂酰甘油和磷脂酸。评估了毒素的抗寄生虫潜力，酸性 PLA 对 T. cruzi 的前鞭毛体形式和 L. infantum 的前鞭毛体形式表现出作用，而碱性 PLA BaspB-II 和 BaspB-IV 对 P. falciparum 表现出活性。结果表明，当 Asp49-PLA 和 Lys49-PLA 相互结合时，抗疟原虫活性增加了 10 倍，表明这些 PLA 同工酶之间存在协同作用。这些发现是首次报道 svPLA 的协同抗疟原虫作用，表明这些分子可能是寻找新的抗寄生虫药物的重要靶标。
