The Transmembrane Protein Center

跨膜蛋白中心(TMPC)建议在PSIBiology网络中发挥协作项目的作用。TMPC研究团队已经组建，在结构基因组学的所有组成任务中提供不同的技能和经验。我们的集体经验表明，需要应用许多技术来扩大成功的机会，因为预计参与PSIBiology Network的膜蛋白的范围很广。该项目已做好充分准备，将提供成功推进PSIBiology网络目标所需的专业知识和领导能力，包括与这一大规模努力的新参与者进行接触。组建的团队包括在膜蛋白的表达、生产和表征方面具有经验的科学家，被证明有能力解决复杂膜蛋白的结构，以及研究膜蛋白与其蛋白质伙伴和生物配体相互作用的良好记录。该团队在生物信息学和生物分析技术的使用方面有很强的基础。每个参与者都带来了膜蛋白功能的独特体验，包括分析开发、制备方法或纯化蛋白和活生物体的研究。这种强调将用于结构确定的膜蛋白的生产与功能表征的能力相结合是我们中心的核心理念。TMPC的主要目标是确定本提案中阐述的每个独特目标类的创建结构。我们的第二个目标是建立一条高效的膜蛋白生产管道，以便逐步提高产量。TMPC希望提高其创造蛋白质变体的能力，根据功能和生物物理特性改进筛选，并改进多个表达途径之间的排序，以找到生产感兴趣蛋白质的最佳方法。强调小规模的净化筛选，并将自动化进一步整合到我们的努力中，将提高效率。结晶筛选的更多选择、同步加速器的使用以及用于解决结构的软件的改进也将有助于提高产量，以实现PSI：生物学目标。

The Transmembrane Protein Center (TMPC) proposes to function as collaborative project within the PSIBiology Network. The TMPC research team has been assembled to provide diverse skills and experience in all constituent tasks of structural genomics. Our collective experience suggests that application of many techniques will be needed to expand the chances for success with the broad range of membrane proteins expected from participation in the PSIBiology Network. This project is well poised to provide the expertise and leadership needed to successfully advance the goals of the PSIBiology Network, including outreach to new participants in this large-scale endeavor. The assembled team includes scientists with experience in the expression, production, and characterization of membrane proteins, proven ability to solve structures of complicated membrane proteins, and a strong track record of studying the interactions of membrane proteins with their protein partners and biological ligands. The team has strong grounding in the use of bioinformatics and bioanalytical techniques. Every participant brings unique experience with the function of membrane proteins, including assay development, preparation methods, or studies with either purified proteins and living organisms. This emphasis on merging the production of membrane proteins for structure determination with the power of functional characterizations is the central philosophy of our Center. The TMPC has a primary goal of determining a founding structure for each of the unique target classes elaborated in this proposal. Our second goal is to establish an efficient membrane protein production pipeline so that increases in throughput may be progressively obtained. The TMPC expects to improve its ability to create protein variants, to refine screening based on functional and biophysical characterizations, and to improve sorting between multiple expressions pathways to find the best way produce a protein of interest. Emphasis on small-scale purification screening and further integration of automation into our efforts will increase efficiency. Expanded options for crystallization screening, access to synchrotrons, and improvements in software used to solve structures will also contribute to the increased throughput necessary to achieve PSI:Biology goals.