Protein Purification and Biochemistry

这一共享资源核心的总体目标是提高项目调查员的能力， 这个合作研究中心，以确定新的分子机制，有助于先天性 免疫识别的各种病毒病原体作出贡献的以下中心目标：1）， 研究新型模式识别受体（PRR）作为病毒核酸真实受体的作用 其影响随后的人类对病毒病原体的先天免疫应答和（2）利用独特的 生物化学能力，以量化技术上具有挑战性的PRR的配体结合功能。 该核心将为中心研究人员提供纯化的先天免疫信号蛋白， 与信号转导相关的生物化学活性的专门测定。芯会 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传导领域的具体工作， 为了提供这种支持。核心将产生这些蛋白质，并测定ATP结合，ATP 水解，并与每种蛋白质的病毒衍生核酸结合，以确保生物化学活性 研究中心研究者提出的研究可获得功能正常的蛋白质。列入该 本U19应用程序中的蛋白质纯化和生物化学核心将为以下方面提供共同来源： 重组信号蛋白，其中几个被用于多个项目。因为生产和 这些蛋白质的表征不是常规的，需要特定的核苷酸测定技术， 结合和水解以及核酸结合，单个共享资源核心将允许高效的 向多个实验室提供这些试剂，并使用可靠的标准化方法， 以评估这些蛋白质的核酸传感功能。总的来说，这个核心将为每个人提供工具， 该项目旨在了解病毒核酸检测的机制，并更好地评估这些机制的潜力。 信号传导系统用于开发基于宿主信号传导途径的新型抗病毒策略。

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.