TRANSIENT CIRCULAR DICHROISM STUDIES OF PROTEIN MOTIONS

时间分辨圆二色谱(TRCD)是一种新的光谱技术 我们在实验室里开发的实验技术 蛋白质构象动力学研究。这份提案提出了 继续和新的生化过程的研究使用TRCD和 互补的时间分辨激光光谱仪。 在赠款期间，与 TRCD技术及其在光谱学研究中的应用 生物分子中的构象运动将被继续研究。它是 我们的目标是开发收集瞬时紫外线的技术 圆二色谱。我们方法的实验细节是 概述。拟议的改善措施将使收集 205 nm至250 nm之间的紫外圆二色光谱 所需的时间延迟介于30皮秒和10微秒之间 光激发。光谱的这一区域监测 蛋白质骨架的吸收。紫外光中的CD光谱 提供了蛋白质中存在的二级结构的度量 解决方案。以下关于血红素蛋白松弛的继续研究 将进行配体的光解离。在这期间的主要努力 下一个资助期将专注于使用UV CD技术以及 用于检查(A)次级的各种激光光谱探头 蛋白质折叠和去折叠过程中的结构演化和(B) 小分子多肽的光诱导螺旋-螺旋转变。蛋白 折叠是生物化学中的一个重要问题。技术的应用 时间分辨紫外圆二色光谱为解决这一问题提供了新的思路 以及对与形成相关的时间尺度的令人兴奋的洞察 蛋白质的二级结构。还将进行有针对性的研究 开发新的光化学方法来触发蛋白质展开 和折叠过程。

Time-resolved circular dichroism (TRCD) spectroscopy is a new experimental technique that we have developed in our laboratory for the study of protein conformational dynamics. This proposal presents continued and new studies of biochemical processes using TRCD and complementary time-resolved laser spectroscopies. During the grant period, both technical developments associated with the TRCD technique and applications of this spectroscopy to the study of conformational motions in biological molecules will be pursued. It is our goal to develop the technology for collecting transient ultraviolet circular dichroism spectra. The experimental details of our approach are outlined. The proposed improvements will enable collection of ultraviolet circular dichroism spectra between 205 nm and 250 nm, at any desired time delay between 30 picoseconds and 10 microseconds following photoexcitation. This region of the optical spectrum monitors the absorption of the protein backbone. CD spectra in the ultraviolet provide a measure of the secondary structures present in proteins in solution. Continued studies on the relaxation of heme proteins following ligand photodissociation will be conducted. The major effort during this next grant period will focus on using the UV CD technique along with a variety of laser spectroscopic probes to examine the (a) secondary structure evolution in the folding and unfolding of proteins and (b) photo-induced helix-coil transitions in small polypeptides. Protein folding is an important problem in biochemistry. The application of time resolved UV CD spectroscopy to this problem offers to produce new and exciting insights into the time scales associated with the formation of protein secondary structure. Research will also be conducted aimed at developing new photochemical methods for triggering protein unfolding and folding processes.