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调查辣木种子产卵刺激素凝集素(WSMoL)与埃及伊蚊腿部存在的膜蛋白结合的能力。

Investigation of the ability of the oviposition-stimulant lectin from Moringa oleifera seeds (WSMoL) to bind with membrane proteins present in the legs of Aedes aegypti.

机构信息

Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, Recife, Brazil.

Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.

出版信息

Int J Biol Macromol. 2020 Nov 1;162:657-662. doi: 10.1016/j.ijbiomac.2020.06.189. Epub 2020 Jun 22.

Abstract

The mosquito Aedes aegypti L. is a vector transmitting diseases such as dengue, chikungunya and Zika virus fever. The water-soluble lectin from Moringa oleifera Lam. seeds (WSMoL) is larvicidal, ovicidal and can stimulate oviposition in A. aegypti. This study aimed to investigate whether WSMoL could bind to membrane proteins from A. aegypti legs. Initially, proteins from the legs were extracted using sodium deoxycholate, digitonin, dodecyl sodium sulfate (SDS) or Triton X-100. The protein concentration was found to be higher in the extract obtained using Triton X-100, which was applied to a WSMoL-Sepharose column. The adsorbed proteins were evaluated using gel filtration chromatography and polyacrylamide gel electrophoresis (PAGE) in presence of SDS. The similarity in the sequences of adsorbed proteins with those available in databases was determined. The proteins adsorbed on the matrix were eluted forming a single peak. Gel filtration chromatography and SDS-PAGE revealed the presence of proteins with molecular masses of approximately 20 kDa and polypeptide bands of 17.0 and 23.7 kDa, respectively. MS/MS analysis indicated similarity between these proteins and ABC carriers, which are expressed in the legs of mosquitos. WSMoL could bind to membrane proteins in the legs of A. aegypti females and induce oviposition through these interactions.

摘要

埃及伊蚊 Aedes aegypti L. 是一种传播登革热、基孔肯雅热和寨卡病毒热等疾病的媒介。辣木 Moringa oleifera Lam. 种子中的水溶性凝集素(WSMoL)具有杀幼虫、杀卵和刺激埃及伊蚊产卵的作用。本研究旨在探讨 WSMoL 是否能与埃及伊蚊腿部的膜蛋白结合。首先,使用脱氧胆酸钠、十二烷基硫酸钠(SDS)或 Triton X-100 从腿部提取蛋白质。结果发现,使用 Triton X-100 提取的蛋白质浓度更高,然后将其应用于 WSMoL-Sepharose 柱。用凝胶过滤层析和 SDS-聚丙烯酰胺凝胶电泳(PAGE)评估吸附的蛋白质。确定与数据库中可用序列相似的吸附蛋白。吸附在基质上的蛋白质形成单个洗脱峰。凝胶过滤层析和 SDS-PAGE 显示存在分子量约为 20 kDa 的蛋白质和分子量分别为 17.0 和 23.7 kDa 的多肽带。MS/MS 分析表明,这些蛋白与 ABC 载体具有相似性,ABC 载体在蚊子的腿部表达。WSMoL 可以与埃及伊蚊雌性腿部的膜蛋白结合,并通过这些相互作用诱导产卵。

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