Catalytic definition of insecticide-metabolizing P450s

杀虫剂代谢 P450 的催化定义

基本信息

批准号：
7097335
负责人：
Mary A Schuler
金额：
$ 29.92万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-01 至 2008-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7097335
关键词：
Anopheles Baculoviridae Drosophilidae X ray crystallography active sites cytochrome P450 drug screening /evaluation enzyme induction /repression enzyme mechanism enzyme substrate high performance liquid chromatography high throughput technology insecticide resistance ligands model design /development molecular dynamics mutant oxidoreductase inhibitor

项目摘要

DESCRIPTION (provided by the applicant): Insects are continually exposed to an array of natural toxic plant chemicals and synthetic toxic insecticides that are used in the management of insects vectoring human and animal diseases and damaging crops. In a growing number of cases, the evolution of P450-mediated detoxification mechanisms have allowed insects to metabolize these compounds and enabled them to survive in the presence of high concentrations of plant toxins and insecticides. From the inception of synthetic insecticide usage in the 1940's, the acquisition of insecticide resistances in insects has increased to the point that they exist in over 500 species with many in species that significantly impact human health, such as the Anopheles mosquitoes that vector malarial parasites (estimated to kill as many as 3 million people per year) and the Aedes and Culex mosquitoes that vector yellow fever and West Nile disease. Our understanding of the molecular basis for these resistances and our ability to counteract these resistances depends on the development of comprehensive molecular models for insect P450s that couple theoretical modeling with protein expression and directed mutagenesis aimed at biochemically testing these models. The projects described are aimed at extending our P450 modeling and protein expression efforts to characterization of the catalytic sites in insecticide-metabolizing P450s in Drosophila, a model organism for the development of insecticide resistance, and in Anopheles, a significant vector for many human pathogens. At the level of molecular modeling, comparisons of these structures can provide information on the similarities (if any) of catalytic sites capable of handling insecticides, the differences that preclude some catalytic sites from binding and metabolizing insecticides and predictions on potential substrates and inhibitors for each protein. At the level of protein expression and high-throughput screening, profiling the range of compounds capable of binding to each catalytic site can define compounds potentially capable of interfering with insecticide metabolisms and, therefore, growth of pathogen-bearing insects resistant to current insecticides. The interdisciplinary mixture of molecular, biochemical and theoretical approaches used in this analysis are well beyond those available to most researchers in the field of insecticide resistance and provide us with significant potential for identifying inhibitors for P450s in mosquito species posing significant health risks as disease vectors.

描述（由申请人提供）：昆虫持续接触一系列天然有毒植物化学品和合成有毒杀虫剂，这些化学物质用于管理传播人类和动物疾病并损害农作物的昆虫。在越来越多的情况下，P450介导的解毒机制的进化使昆虫能够代谢这些化合物，并使它们能够在高浓度的植物毒素和杀虫剂存在下生存。自 20 世纪 40 年代开始使用合成杀虫剂以来，昆虫对杀虫剂产生的抗药性已经增加到 500 多种昆虫中都存在这种抗药性，其中许多昆虫对人类健康有重大影响，例如传播疟疾寄生虫的按蚊。据估计每年导致多达 300 万人死亡）以及传播黄热病和西尼罗河病的伊蚊和库蚊。我们对这些抗性的分子基础的理解以及我们对抗这些抗性的能力取决于昆虫 P450 综合分子模型的开发，该模型将理论模型与蛋白质表达和旨在生化测试这些模型的定向诱变相结合。所描述的项目旨在扩展我们的 P450 建模和蛋白质表达工作，以表征果蝇（一种产生杀虫剂抗性的模式生物）和按蚊（许多人类病原体的重要载体）中杀虫剂代谢 P450 的催化位点。在分子建模水平上，这些结构的比较可以提供有关能够处理杀虫剂的催化位点的相似性（如果有的话）、阻止某些催化位点结合和代谢杀虫剂的差异以及对每种催化位点的潜在底物和抑制剂的预测的信息。蛋白质。在蛋白质表达和高通量筛选水平上，分析能够与每个催化位点结合的化合物范围可以确定可能干扰杀虫剂代谢的化合物，从而干扰对当前杀虫剂具有抗性的携带病原体的昆虫的生长。该分析中使用的分子、生物化学和理论方法的跨学科组合远远超出了杀虫剂抗药性领域大多数研究人员可以使用的方法，并为我们在作为疾病媒介构成重大健康风险的蚊子物种中识别 P450 抑制剂提供了巨大的潜力。