STUDYING TRPZIPS USING RESONANCE RAMAN SPECTROSCOPY

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。 β-发夹的折叠动力学一直是GAI小组的一个重要研究领域。我们用静态红外光谱、圆二色谱和红外温度跳跃法研究了色氨酸拉链等一系列β-发夹的结构稳定性和折叠动力学。为了进一步扩大我们对这一系统的了解，我们计划与Spiros博士的实验室合作，在时间分辨模式下使用共振拉曼光谱来研究色氨酸拉链(Trpzips)的运动动力学。色氨酸是一种芳香族残基，被认为参与了氢键相互作用，因此可以用作蛋白质中不同位置运动的探针。因此，Trpzips是一种理想的共振拉曼光谱系统，因为我们可以使用波长选择的激光激发来监测色氨酸侧链的共振拉曼光谱，从而监测Trpzips的运动。我们计划研究Trpzips中在229 nm波长和200 nm波长以下激发的酰胺带中的Trptophan残基。本研究中使用的多肽序列如下：Trpzip2：SWTWENGKWTWK。作为初步研究，我们想做一组静态测量和1到5组T跳跃测量，以检查激活能。如果我们获得了有利的结果，我们将进一步扩大研究范围。

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. Folding dynamics of beta-hairpins has been a prominent area of studies in the Gai group. We have investigated the structural stability and folding kinetics of a series of beta-hairpins, namely: tryptophan zippers, by static Infrared spectroscopy, Circular Dichroism and the Infrared temperature jump method. To further extend our understanding of this system, we plan to collaborate with Dr. Spiros lab and use Resonance Raman Spectroscopy in a time resolved mode to study the dynamics of motion of tryptophan zippers (Trpzips). Tryptophan is an aromatic residue and is believed to be involved in H- bonding interactions, and thus can be used as probes of motion at various sites in the protein. Therefore, Trpzips are an ideal system for Resonance Raman Spectroscopy because we can use wavelength-selective laser excitation to monitor Resonance Raman spectra of the tryptophan sidechains and thus monitor the motion of Trpzips. We plan look at investigate Trptophan residues in the Trpzips at 229 nm wavelength and amide bands excited below 200 nm wavelength. The sequence of the peptide used in the current study is as following: Trpzip2: SWTWENGKWTWK. For preliminary studies, we would like to do one set of static measurement and 1 to 5 sets of T-jump measurements so as to check the activation energy. If we obtain favorable results we will extend the study further.