Characterization of ABC transporters in parasitic nematodes

寄生线虫 ABC 转运蛋白的表征

• 批准号: 293164-2007
• 负责人: Ardelli, Bernadette
• 金额: $ 1.77万
• 依托单位: Brandon University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2014
• 资助国家: 加拿大
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=548201
• 关键词: Characterization; ABC; transporters; parasitic; nematodes

ABC transporters have been found in all living organisms and form one of the largest characterized protein families. Organisms that live in diverse environments and that must adapt to various external conditions have a large repertoire of ABC transporters (e.g., Caenorhabditis elegans chromosomes encode 60 ABC transporters) while organisms that occupy a more restricted growth niche have considerably less (e.g., Trypanosoma brucei chromosomes encode 21 ABC transporters). ABC transporters are integral to many cellular processes, yet little is known of these proteins in parasitic nematodes. In farm animals and humans, nematodes cause disease, death and economic loss. Essential to their survival are transport mechanisms, which include ABC transporters. All anthelmintics currently used to treat nematodes target transporters. Some ABC transporter gene sequences have been identified in parasitic nematodes however, functional studies do not exist. The long-term goal of the research is to understand ABC transporter mediated drug resistance in nematodes. Before this can be accomplished, the basic knowledge of ABC transporters in parasitic nematodes must be expanded. To do this Brugia malayi, a nematode that resides in the lymphatic system of humans, will be used as the model organism. The specific goals of the research are to (1) use bioinformatic and genomic analyses to identify and classify the Brugia ABC transporter superfamily; (2) create a spatial proteome of Brugia transporters by transfection of C. elegans with parasite genes; (3) determine the expression patterns in all sexual and life cycle stages using quantitative RT-PCR; and (4) determine the substrate specificity and transport kinetics of Brugia transporters using site-directed mutagenesis and binding assays. The research has implications for expanding the basic knowledge of ABC transporters in parasitic nematodes and providing insight into their cellular functions. As many ABC transporters are targets for anthelmintics, this research will suggest new or alternative control strategies and also expand the existing knowledge of the mechanism of resistance to anthelmintics.

ABC转运蛋白存在于所有生物体中，是最大的蛋白家族之一。生活在不同环境中并且必须适应各种外部条件的生物体拥有大量的ABC转运蛋白（例如秀丽隐杆线虫染色体编码60个ABC转运蛋白），而占据更有限生长生态位的生物体则拥有相当少的ABC转运蛋白（例如，布鲁氏锥虫染色体编码21个ABC转运蛋白）。ABC转运蛋白是许多细胞过程的组成部分，但对寄生线虫中的这些蛋白知之甚少。在农场动物和人类中，线虫会导致疾病、死亡和经济损失。它们生存的关键是转运机制，其中包括ABC转运蛋白。目前用于治疗线虫的所有驱虫剂都以转运体为目标。一些ABC转运蛋白基因序列已在寄生线虫中被鉴定出来，但功能研究尚不存在。该研究的长期目标是了解ABC转运蛋白介导的线虫耐药。在此之前，必须扩大对寄生线虫ABC转运蛋白的基本知识。为了做到这一点，一种居住在人类淋巴系统中的线虫将被用作模式生物。该研究的具体目标是：(1)使用生物信息学和基因组学分析来识别和分类Brugia ABC转运蛋白超家族；(2)用疟原虫基因转染秀丽隐杆线虫，构建Brugia转运体空间蛋白质组；(3)利用定量RT-PCR技术确定各有性和生命周期阶段的表达模式；(4)利用定点诱变和结合试验确定布鲁吉亚转运蛋白的底物特异性和转运动力学。该研究对扩大寄生线虫ABC转运蛋白的基础知识和深入了解其细胞功能具有重要意义。由于许多ABC转运蛋白是驱虫药的靶点，本研究将提出新的或替代的控制策略，并扩展对驱虫药抗性机制的现有知识。