LARGE-SCALE MEASUREMENT OF PROTEIN THERMODYNAMIC PARAMETERS

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。 测量蛋白质响应于增加浓度的化学变性剂的解折叠可用于推导其热力学稳定性。 T7噬菌体在相对高浓度的变性剂中是稳定的。 我们已经利用这一事实进行变性实验的突变体的WW结构域的大规模。 我们将噬菌体展示的WW结构域变体的文库暴露于增加浓度的化学变性剂胍，然后进行肽结合的选择。 通过检查每种变体在增加浓度的胍中的性能，我们可以计算每种变体的相对热力学稳定性。 由此产生的WW域的稳定性地图应该提供一个前所未有的视图区域管理这个域的稳定性，并可能阐明蛋白质稳定性的一般原则。

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. Measuring the unfolding of a protein in response to increasing concentrations of chemical denaturants can be used to derive its thermodynamic stability. T7 phage are stable in relatively high concentrations of denaturant. We have exploited this fact to perform denaturation experiments on mutants of a WW domain on a massive scale. We exposed a library of phage-displayed WW domain variants to increasing concentrations of the chemical denaturant guanidine and then performed a selection for peptide binding. By examining the performance of each variant in increasing concentrations of guanidine, we can calculate the relative thermodynamic stability of each variant. The resulting stability map of the WW domain should offer an unprecedented view of regions governing the stability of this domain and may illuminate general principles of protein stability.