LARGE SCALE MEASUREMENT OF EPISTASIS TO IDENTIFY MUTATIONS THAT STABILIZE PROTEI

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 工程化蛋白质以增强稳定性对于它们的工业或制药用途通常是至关重要的，但是实现这一目标的实验和计算努力已经取得了有限的成功。 我们仅基于对超过47，000个变体的蛋白质适应性的测量来确定WW结构域中的稳定突变，并使用这些适应性测量来计算超过5，000个上位关系。 我们引入了一个新的指标，称为？合作伙伴增强，？其是当在双突变体中组合时突变赋予其它突变的平均上位效应的量度。 该度量允许我们推断WW结构域内的15个稳定突变的身份。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Engineering proteins for enhanced stability is often critical for their industrial or pharmaceutical use, but experimental and computational efforts to accomplish this goal have met with limited success. We identified stabilizing mutations in a WW domain based solely on a measurement of protein fitness for more than 47,000 variants, and the use of these fitness measurements to calculate more than 5,000 epistastic relationships. We introduce a new metric, termed ?partner potentiation,? which is a measure of the mean epistasis effect that a mutation imparts to other mutations when combined in a double mutant. This metric allowed us to infer the identity of 15 stabilizing mutations within the WW domain.