RUI: Collaborative Research: Head Group Preference in Recluse Spider Phospholipase D Toxins

RUI：合作研究：隐士蜘蛛磷脂酶 D 毒素的头群偏好

• 批准号: 1807885
• 负责人: Greta Binford
• 金额: $ 12万
• 依托单位: Lewis and Clark College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1807885&HistoricalAwards=false
• 关键词: RUI; Collaborative; Research; Head; Group

With this award, the Chemistry of Life Processes Program in the Chemistry Division is supporting a collaborative project between Dr. Matthew Cordes at the University of Arizona and Dr. Greta Binford at Lewis and Clark College, to determine how spider venom toxins target and destroy different molecules on cell surfaces. Venoms of brown recluse spiders have toxins that cause the death of cells and tissue in mammals, but also help the spiders immobilize and/or digest insect prey. The toxins that are known to affect mammals can damage cell surfaces by cutting a specific "head group", called choline, off of a molecule called a sphingolipid. Other recluse spider toxins can only cut off a different kind of head group, called ethanolamine, that mammals do not have in their sphingolipids, so these toxins may be less harmful to mammals. Many insects have both kinds of head group, so both types of toxin are probably important for predation, perhaps in different ways. Dr. Cordes, a structural biologist and biochemist, and Dr. Binford, a biologist and expert on venomous invertebrates, combine their expertise to determine how these toxins distinguish the different head groups, and what the biological consequences are for predators and their prey. The specific action of these toxins on different head groups could also make them useful in biotechnology for detecting or manipulating different kinds of cell surfaces. The broader impacts of this project involve outreach through mentorship of undergraduates doing integrative and collaborative research at two different institutions. Spiders attract public interest and afford an opportunity to inform the public on scientific investigations on the subject. An innovative program known as "SpiderFest" is conduucted both at the laboratory and at a science expo in Portland. The overall goal of this project is to understand the causes and effects of substrate head group (ethanolamine vs. choline) preference in phospholipase D toxins from recluse spiders. The specific goals are to map interfacial binding sites in the toxins, elucidate amino-acid sequence determinants of substrate head group preference; characterize the evolution of substrate preference in the recluse spider toxin family; and correlate head group preference to biological effects of the toxins. Methods to be used include NMR, X-ray crystallography, computational structural biology, site-directed mutagenesis, phylogenetic reconstruction, enzymatic assays, and biological assays. Information from this study illuminates differential recognition, by proteins that act at membrane surfaces, of the two most common lipid head groups in nature, phosphocholine and phosphoethanolamine. The project also sheds light on toxin recruitment and specialization in venoms and interesting aspects of arthropod biochemistry and neurobiology. These toxins could also be developed into valuable analytical tools to probe important differences in cell surface structure.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

有了这个奖项，化学生命过程的化学过程计划支持亚利桑那大学的Matthew Cordes博士与Lewis和Clark College的Greta Binford博士之间的合作项目，以确定蜘蛛毒素的目标并破坏细胞表面上的不同分子。 棕色隐居蜘蛛的毒液具有导致哺乳动物细胞和组织死亡的毒素，但也有助于蜘蛛固定和/或消化昆虫猎物。 已知会影响哺乳动物的毒素会通过切开称为胆碱的分子的特定“头部组”（称为胆碱）来损害细胞表面。其他隐居蜘蛛毒素只能切断哺乳动物没有鞘脂的另一种称为乙醇胺的头部组，因此这些毒素可能对哺乳动物有害。许多昆虫都有两种头群，因此两种类型的毒素可能对捕食很重要，也许以不同的方式。结构性生物学家和生物化学家Cordes博士以及有毒无脊椎动物的生物学家兼专家Binford博士结合了其专业知识，以确定这些毒素如何区分不同的头部组，以及对捕食者及其猎物的生物学后果是什么。这些毒素在不同头部组上的具体作用也可能使它们在检测或操纵各种细胞表面的生物技术中有用。 该项目的更广泛影响涉及通过在两个不同机构进行综合和协作研究的大学生的指导来推广。 蜘蛛吸引了公众的兴趣，并有机会向公众提供有关该主题的科学研究的机会。在实验室和波特兰的一家科学博览会上，都纵容了一个被称为“蜘蛛网”的创新计划。该项目的总体目标是了解磷脂酶D毒素中底物头组（乙醇胺与胆碱）偏好的原因和影响。具体目标是绘制毒素中的界面结合位点，阐明底物头组偏好的氨基酸序列决定因素。表征了隐居蜘蛛毒素家族中底物偏好的演变；并将头部组的偏好与毒素的生物学作用相关。要使用的方法包括NMR，X射线晶体学，计算结构生物学，定向诱变，系统发育重建，酶试验和生物学测定。这项研究的信息通过作用于膜表面的蛋白质，在自然界中两个最常见的脂质头群，磷酸胆碱和磷酸乙醇胺的蛋白质来阐明差异识别。该项目还阐明了毒素的招募和专业化，并在节肢动物生物化学和神经生物学的有趣方面介绍了毒素。这些毒素也可以发展为有价值的分析工具，以探测细胞表面结构的重要差异。该奖项反映了NSF的法定任务，并被认为是值得通过基金会的知识分子优点和更广泛的影响评估标准通过评估来支持的。