Integrated Center for Structure and Function Innovation

结构与功能创新综合中心

• 批准号: 7088975
• 负责人: THOMAS C. TERWILLIGER
• 金额: $ 409.02万
• 依托单位: LOS ALAMOS NAT SECTY-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7088975
• 关键词: biomedical resource; cooperative study; functional /structural genomics; protein structure function; proteomics

DESCRIPTION (provided by applicant): The Integrated Center for Structure and Function Innovation (ISFI) will be a PSI-2 Specialized Center developing and applying a set of synergistic technologies organized to overcome recognized bottlenecks in structure determination at the key steps of production of soluble protein and protein crystallization. The development of these technologies will be embedded in an integrated pipeline for protein structure determination and analysis of function, and the technologies will be applied to difficult proteins and protein complexes from large sequence families, emphasizing proteins with high biological significance. The increased success rates made possible by our technologies will be critical for obtaining structures of proteins and protein complexes of high functional value, both for the PSI and for structural biology as a whole. The increased success rates will lead as well to substantial cost reductions in structure determination. Our application of our methodologies as part of this project will enhance the understanding of protein function and protein-protein interactions relevant to human health. The core technologies to be developed in the ISFI are: (1) systematic construction of protein binding partners for chaperone-assisted crystallography, (2) engineering proteins for crystallization by surface mutagenesis and addition of fusion proteins, (3) directed evolution of proteins for solubility and trapping of protein partners required for folding using one member attached to a reporter protein as a bait, (4) identification of protein partners required for proper folding using informatics approaches confirmed by experiments, and (5) systematic combinatorial crystallization screening and optimization. These technologies will be used to address five major bottlenecks in the structural genomics pipeline: (a) soluble expression, (b) small protein complexes, (c) crystallizability, (d) soluble domains of membrane proteins, and (e) eukaryotic protein structure determination. The technologies will be developed through highly integrated collaborations among 6 institutions. Technologies and results will be made available to other Centers and the broader structural biology community through workshops, publications, personnel exchanges, and the web.

描述（由申请人提供）：结构和功能创新综合中心（ISFI）将是一个PSI-2专业中心，开发和应用一套协同技术，以克服可溶性蛋白质生产和蛋白质结晶关键步骤中结构测定的公认瓶颈。 这些技术的开发将嵌入蛋白质结构测定和功能分析的集成管道中，这些技术将应用于来自大序列家族的困难蛋白质和蛋白质复合物，强调具有高生物学意义的蛋白质。 通过我们的技术提高成功率对于获得具有高功能价值的蛋白质和蛋白质复合物的结构至关重要，无论是对于PSI还是对于整个结构生物学。成功率的提高也将导致结构确定的成本大幅降低。 作为该项目的一部分，我们的方法学应用将增强对与人类健康相关的蛋白质功能和蛋白质-蛋白质相互作用的理解。 ISFI将开发的核心技术是：（1）用于分子伴侣辅助结晶学的蛋白质结合配偶体的系统构建，（2）通过表面诱变和加入融合蛋白来工程化用于结晶的蛋白质，（3）使用连接到报告蛋白的一个成员作为诱饵，针对溶解性和捕获折叠所需的蛋白质配偶体的蛋白质定向进化，（4）使用经实验证实的信息学方法鉴定正确折叠所需的蛋白质配偶体，以及（5）系统的组合结晶筛选和优化。 这些技术将用于解决结构基因组学管道中的五个主要瓶颈：（a）可溶性表达，（B）小蛋白复合物，（c）结晶性，（d）膜蛋白的可溶性结构域，以及（e）真核蛋白结构测定。 这些技术将通过6个机构之间的高度集成合作来开发。技术和成果将通过研讨会、出版物、人员交流和网络提供给其他中心和更广泛的结构生物学社区。