Bothrops atrox venom: Biochemical properties and cellular phenotypes of three highly toxic classes of toxins
Bothrops atrox venom: Biochemical properties and cellular phenotypes of three highly toxic classes of toxins
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DOI：
10.1016/j.bbapap.2023.140930
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发表时间：
2023-07-25
影响因子：
3.2
通讯作者：
Chavez-Olortegul, Carlos
中科院分区：
文献类型：
--
作者：
Lopes-de-Souza, Leticia;Costal-Oliveira, Fernanda;Chavez-Olortegul, Carlos
蛇毒是一种复杂的化合物混合物，在不同物种之间是保守的，并具有协同作用，引发严重的局部和全身影响。确定对细胞动态平衡最具破坏性的毒素类别，将是专注于支撑毒液的主要活动的有效方法。在这里，我们将重点介绍南美洲亚马逊地区发生的大部分事故的罪魁祸首--黑斑蛇的毒液。我们鉴定了阿曲霉毒素的关键细胞毒素组分，并绘制了它们的生化性质、蛋白质组成和细胞损伤的图谱。通过质量排除层析得到五个组分，它们要么含有单一的酶活性(即L-氨基酸氧化酶或透明质酸酶)，要么含有两个或两个以上蛋白质组分的不同活性(如金属蛋白酶、丝氨酸蛋白酶或磷脂酶A2)。只有三种蛋白质组分降低了原代人类细胞的活力。引人注目的是，这样的活动伴随着细胞与底物和邻近细胞的附着中断。这种形态细胞特征的强烈扰动表明活体组织完整性可能存在缺陷。质谱仪确定了导致这些表型的主要毒素类别。在这里，我们提供了一种选择关键细胞毒蛋白的策略，用于有针对性地研究它们在蛇咬伤毒化过程中的作用机制和潜在的协同作用。我们的数据强调了推测的毒素(或其组合)，这可能是未来治疗干预的重点。
Snake venoms have a complex mixture of compounds that are conserved across species and act synergistically, triggering severe local and systemic effects. Identification of the toxin classes that are most damaging to cell homeostasis would be a powerful approach to focus on the main activities that underpin envenomation. Here, we focus on the venom of Bothrops atrox, snake responsible for most of the accidents in Amazon region of South America. We identified the key cytotoxic toxin fractions from B. atrox venom and mapped their biochemical properties, protein composition and cell damage. Five fractions were obtained by mass exclusion chromatography and contained either a single class of enzymatic activity (i.e., L-amino acid oxidases or Hyaluronidases) or different activities co-distributed in two or more protein fractions (e.g., Metalloproteinases, Serine Proteases, or Phospholipases A2). Only three protein fractions reduced cell viability of primary human cells. Strikingly, such activity is accompanied by disruption of cell attachment to substratum and to neighbouring cells. Such strong perturbation of morphological cell features indicates likely defects in tissue integrity in vivo. Mass spectrometry identified the main classes of toxins that contribute to these phenotypes. We provide here a strategy for the selection of key cytotoxic proteins for targeted investigation of their mechanism of action and potential synergism during snakebite envenomation. Our data highlights putative toxins (or combinations of) that may be the focus of future therapeutic interference.