LARGE SCALE MEASUREMENT OF EPISTASIS TO IDENTIFY MUTATIONS THAT STABILIZE PROTEI

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 设计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，但实现这一目标的实验和计算努力取得的成功有限。我们仅根据对47,000多个变体的蛋白质适合度的测量，并使用这些适合度测量来计算超过5,000个上位关系，从而确定了WW结构域中的稳定突变。我们引入了一个新的度量，称为？伴侣增强，？这是一种衡量一个突变赋予其他突变的平均上位性效应的指标，当组合在一个双突变中时。这一指标使我们能够推断出WW结构域中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.