Protein Purification and Biochemistry

这一共享资源核心的总体目标是增强项目调查员的能力 这个合作研究中心确定了先天致病的新分子机制 对各种病毒病原体的免疫识别有助于实现以下中心目标：1) 探讨新型模式识别受体(PRR)作为病毒核酸真实受体的作用 它们影响随后的人类对病毒病原体的先天免疫反应，以及(2)利用独特的 量化PRRs的配体结合功能的生化能力，这在技术上具有挑战性。 该核心将为中心调查人员提供纯化的天然免疫信号蛋白以及 与信号转导相关的生化活动的专门分析。核心意志 利用邓肯博士在重组蛋白生产和表征方面的专业知识， 信号通路的生化重建，以及在先天免疫信号领域的具体工作 才能提供这种支持。核心将产生这些蛋白质，并检测与ATP结合的ATP 水解，并与病毒衍生的每种蛋白质的核酸结合，以确保生物化学活性 对于中心研究人员提出的研究，功能正常的蛋白质是可用的。包括 U19应用中的蛋白质纯化和生化核心将提供一个共同的来源 重组信号蛋白，其中几个被用于多个项目。因为它的制作和制作 这些蛋白质的特性不是常规的，需要特定的核苷酸分析技术 结合和水解以及核酸结合，单一的共享资源核心将允许高效 将这些试剂提供给多个实验室，并按顺序使用强大的标准化方法 以评估这些蛋白质的核酸传感功能。总体而言，该核心将为每个 了解病毒核酸检测机制并更好地评估其潜力的项目 信号系统将用于开发基于宿主信号通路的新的抗病毒策略。

The overall objective of this Shared Resource Core is to enhance the capacity of the project investigators of this cooperative research center to determine the novel molecular mechanisms contributing to innate immune recognition of a variety of viral pathogens by contributing to the following center goals: 1) to investigate the role novel Pattern Recognition Receptors (PRR) as authentic receptors of viral nucleic acid which affect subsequent human innate immune responses to viral pathogens and (2) to capitalize on unique biochemical capabilities to quantify the ligand-binding functions of PRRs that are technically-challenging. The core will provide center investigators with purified innate immune signaling proteins as well as specialized assays of their biochemical activities that are relevant to signal transduction. The Core will capitalize on the expertise of Dr. Duncan in recombinant protein production and characterization, biochemical reconstitution of signaling pathways, and specific work in the field of innate immune signaling in order to provide this support. The core will generate these proteins and assay the ATP binding, ATP hydrolysis, and binding to virally derived nucleic acids of each protein in order to ensure biochemically active properly functioning protein is available for the studies proposed by center investigators. The inclusion of the Protein Purification and Biochemistry Core in this U19 application will provide a common source for recombinant signaling proteins, several of which are utilized in multiple projects. Because the production and characterization of these proteins is not routine and requires specific assay technologies for nucleotide binding and hydrolysis as well as nucleic acid binding, a single Shared resource Core will allow for efficient availability these reagents to multiple laboratories and the use of robust standardized methodologies in order to assess nucleic acid sensing functions by these proteins. Overall, this core will provide tools for each project to understand the mechanisms of viral nucleic acid detection and better assess the potential for these signaling systems to be used to develop novel anti-viral strategies based on host signaling pathways.