Catalytic definition of insecticide-metabolizing P450s

杀虫剂代谢 P450 的催化定义

• 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
• 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

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）：昆虫持续暴露于一系列天然有毒植物化学品和合成有毒杀虫剂，这些化学品和合成有毒杀虫剂用于管理传播人类和动物疾病并破坏作物的昆虫。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植物毒素和杀虫剂的存在下生存。从20世纪40年代开始使用合成杀虫剂开始，昆虫中杀虫剂抗性的获得已经增加到它们存在于500多个物种中的程度，其中许多物种显著影响人类健康，例如传播疟疾寄生虫的按蚊（估计每年杀死多达300万人）以及携带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和我们对抗这些抗性的能力取决于昆虫P450的综合分子模型的发展，该模型将理论建模与蛋白质表达和定向诱变结合起来，旨在对这些模型进行生化测试。所描述的项目旨在扩展我们的P450建模和蛋白质表达的努力，以表征在果蝇，杀虫剂抗性发展的模式生物，和在按蚊，许多人类病原体的重要载体中的P450代谢的催化位点。在分子建模的水平上，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）的信息，排除一些催化位点结合和代谢杀虫剂的差异，以及对每种蛋白质的潜在底物和抑制剂的预测。在蛋白质表达和高通量筛选的水平上，分析能够与每个催化位点结合的化合物的范围可以定义潜在地能够干扰杀虫剂代谢的化合物，并且因此干扰对当前杀虫剂具有抗性的携带病原体的昆虫的生长。在这项分析中使用的分子，生物化学和理论方法的跨学科的混合物远远超出了大多数研究人员在杀虫剂耐药性领域，并为我们提供了显着的潜力，确定抑制剂P450蚊子物种构成重大的健康风险的疾病载体。