A FRET-BASED SCREEN FOR THE BINDING OF COMPUTATIONALLY DESIGNED PROTEIN PAIRS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 计算生物学的一个目标是有能力设计新的蛋白质伙伴。通过这种方式，可以创造出新的蛋白质活性，这些活性既可以用于基础研究，也可以用于创造人类疾病的新诊断或治疗方法。现在，在开发的早期阶段，这个过程是从两个相互不结合的蛋白质开始，然后重新设计它们，以创建一个特定的、高亲和力的共享结合表面。我们已经开始应用这种方法来设计一对蛋白质，它们通过一种新的蛋白质-蛋白质界面相互作用。由于这个界面完全是计算机设计的产物，它与组成酵母蛋白质组的所有蛋白质完全正交。此功能建议将有趣的应用程序作为一组亲和性标记。 这一过程中的一个限制步骤是评估变化是否改善了结合亲和力。该项目旨在结合YRC在蛋白质设计和FRET方面的专业知识，开发一种快速筛选重新设计的蛋白质形成异二聚体的能力的方法。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. One goal of computational biology is to have the ability to design novel protein partners. In this way novel protein activities could be created that may find use in both basic research, and in creating novel diagnostics or treatments for human disease. Now in an early phase of development, the process is to begin with two proteins that do not bind to each other, and to redesign them to create a specific, high affinity shared binding surface. We have begun by applying this methodology to design a pair of proteins, which interact via a novel protein-protein interface. Since this interface is solely the product of computational design, it is completely orthogonal to all proteins which comprise the yeast proteome. This feature suggests an interesting application as a set of affinity tags. A limiting step in the process is the assessment of whether changes have improved binding affinities. This project aims to combine the YRC expertise in both protein design and FRET to develop a method for rapid screening of the redesigned proteins for their ability to form heterodimers.