Integrated Center for Structure and Function Innovation

描述(由申请人提供)：结构和功能创新综合中心(ISFI)将是PSI-2专业中心，开发和应用一套协同技术，旨在克服在生产可溶性蛋白质和蛋白质结晶的关键步骤中结构确定方面的公认瓶颈。 这些技术的发展将嵌入到蛋白质结构测定和功能分析的综合管道中，这些技术将应用于困难的蛋白质和来自大序列家族的蛋白质复合体，强调具有高度生物学意义的蛋白质。 我们的技术带来的更高的成功率对于获得蛋白质结构和具有高功能价值的蛋白质复合体至关重要，无论是对PSI还是整个结构生物学来说都是如此。成功率的提高还将导致结构确定方面的成本大幅降低。作为该项目的一部分，我们应用我们的方法将加强对蛋白质功能和与人类健康相关的蛋白质-蛋白质相互作用的理解。 将在ISFI中开发的核心技术是：(1)系统地构建用于伴侣辅助结晶学的蛋白质结合伙伴，(2)通过表面突变和添加融合蛋白来设计结晶蛋白质，(3)利用附着在报告蛋白上的一个成员作为诱饵来定向进化蛋白质的溶解性和捕获折叠所需的蛋白质伙伴，(4)利用实验证实的信息学方法确定正确折叠所需的蛋白质伙伴，以及(5)系统的组合结晶筛选和优化。 这些技术将被用来解决结构基因组学管道中的五个主要瓶颈：(A)可溶表达，(B)小蛋白复合体，(C)结晶性，(D)膜蛋白的可溶性域，和(E)真核蛋白质结构测定。 这些技术将通过6个机构之间的高度整合合作来开发。技术和成果将通过研讨会、出版物、人员交流和网络向其他中心和更广泛的结构生物学社区提供。

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.