FAST KINETICS OF DESIGNED SERPINS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 这是一个旨在了解蛋白质如何使用实验和计算技术折叠的项目。后者可以指导快速文件夹和结构一致的突变体的搜索。我们将采用的计算技术使用统计热力学来识别蛋白质序列中的挫折位置以及结构一致和较少挫折的突变。 此方法以前曾用于指导快速文件夹的搜索。 在这种情况下，它已被应用于20个残基的微蛋白，色氨酸笼，在努力减少其4微秒的折叠时间进一步。 我们将使用的实验技术包括激光诱导温度跳跃动力学实验（红外或紫外检测）以及静态实验，使用荧光，红外光谱和圆二色性。 我们进一步建议使用计算辅助肽设计与荧光相关光谱（FCS）和荧光共振能量转移（FRET）为基础的单分子技术来设计和表征短肽，也阻碍和逆转丝氨酸蛋白酶抑制剂的聚合。具体目标是从理论和实验的角度设计和研究这些肽，并研究解聚的基本机制细节，从而为潜在治疗分子的设计提供信息。

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. This is a project aimed to understand how proteins fold using both experimental and computational techniques. The latter can guide the search for fast folders and also tructurally consistent mutants. The computational technique we will employ uses statiscal thermodynamics to identify frustrated positions in the protein sequence as well as structurally consistent and less frustrating mutations. This method has been used previously to guide the search for fast folders. In this case, it has been applied to the 20 residue miniprotein, Trp-cage, in an effort to decrease its 4 microsecond folding time further. The experimental techniques we will use include laser induced temperature-jump kinetics experiments (with IR or UV detection) as well as static experiments using fluorescence, infrared spectroscopy, and circular dichroism. We further propose to use computationally assisted peptide design in association with fluorescence correlation spectroscopy (FCS) and fluorescence resonance energy transfer (FRET) based single molecule techniques to design and characterize short peptides that also impede and reverse the polymerization of serpins. The specific aims are to design and study these peptides both from the theoretical and experimental points of view and to investigate the underlying mechanistic details of depolymerization, thus informing the design of potential therapeutic molecules.