Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps
Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps
复制标题
DOI：
10.3390/toxins11080448
复制
发表时间：
2019-08-01
期刊：
影响因子：
4.2
通讯作者：
de Brandao Prieto-da-Silva, Alvaro Rossan
中科院分区：
文献类型：
--
作者：
Ceolin Mariano, Douglas Oscar;de Oliveira, Ursula Castro;de Brandao Prieto-da-Silva, Alvaro Rossan
蚂蚁物种具有专门的毒液系统，用于叮咬和消化有机化合物的生物鸡尾酒，包括肽和多肽毒素，用于捕食和防御。袋蚁属由捕食性巨蚁组成，其毒液能够引起持久的局部疼痛、不自主的颤抖、淋巴结病和心律失常等症状。为了加深我们对毒液中蛋白质毒素组成及其在化学生态关系和人类健康中的作用的了解，我们对巨型蚂蚁D的粗毒液进行了自下而上的蛋白质组学分析。股四头肌，俗称“假”tocandiras。为此，我们使用了两种不同的分析方法：（i）基于凝胶的蛋白质组学方法，其中通过变性十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）解析粗毒液，并切下所有蛋白条带用于分析;（ii）基于溶液的蛋白质组学方法，其中粗毒液蛋白组分在溶液中直接片段化为胰蛋白酶肽用于分析。将这两种方法得到的蛋白质组数据与先前注释的D.然后，对所有鉴定的转录产物进行同源性搜索。基于凝胶的蛋白质组学方法明确地鉴定了毒液中的9种高分子量毒素，例如，酶[透明质酸酶、磷脂酶A1、二肽基肽酶和葡萄糖脱氢酶/黄素腺嘌呤二核苷酸（FAD）醌]和多种毒液过敏原（与红火蚁Selenopsis invicta同源）和毒液相关蛋白（主要的皇家毒液样蛋白）。此外，基于溶液的蛋白质组学揭示并证实了几种水解酶，氧化还原酶，蛋白酶，Kunitz样多肽，以及丰度较低的抑制剂半胱氨酸结（ICK）样（结素）神经毒素和昆虫防御素的存在。结果表明，D.股四头肌毒是一种具有高度细胞膜和组织损伤、神经毒性和过敏反应的毒素，因此，扩大了对股四头肌毒的认识。四头肌毒液的组成及其对猎物和受害者的潜在生物效应。
Ant species have specialized venom systems developed to sting and inoculate a biological cocktail of organic compounds, including peptide and polypeptide toxins, for the purpose of predation and defense. The genus Dinoponera comprises predatory giant ants that inoculate venom capable of causing long-lasting local pain, involuntary shaking, lymphadenopathy, and cardiac arrhythmias, among other symptoms. To deepen our knowledge about venom composition with regard to protein toxins and their roles in the chemical-ecological relationship and human health, we performed a bottom-up proteomics analysis of the crude venom of the giant ant D. quadriceps, popularly known as the "false" tocandiras. For this purpose, we used two different analytical approaches: (i) gel-based proteomics approach, wherein the crude venom was resolved by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and all protein bands were excised for analysis; (ii) solution-based proteomics approach, wherein the crude venom protein components were directly fragmented into tryptic peptides in solution for analysis. The proteomic data that resulted from these two methodologies were compared against a previously annotated transcriptomic database of D. quadriceps, and subsequently, a homology search was performed for all identified transcript products. The gel-based proteomics approach unequivocally identified nine toxins of high molecular mass in the venom, as for example, enzymes [hyaluronidase, phospholipase A1, dipeptidyl peptidase and glucose dehydrogenase/flavin adenine dinucleotide (FAD) quinone] and diverse venom allergens (homologous of the red fire ant Selenopsis invicta) and venom-related proteins (major royal jelly-like). Moreover, the solution-based proteomics revealed and confirmed the presence of several hydrolases, oxidoreductases, proteases, Kunitz-like polypeptides, and the less abundant inhibitor cysteine knot (ICK)-like (knottin) neurotoxins and insect defensin. Our results showed that the major components of the D. quadriceps venom are toxins that are highly likely to damage cell membranes and tissue, to cause neurotoxicity, and to induce allergic reactions, thus, expanding the knowledge about D. quadriceps venom composition and its potential biological effects on prey and victims.