TRANSIENT CIRCULAR DICHROISM STUDIES OF PROTEIN MOTIONS

蛋白质运动的瞬态圆二色性研究

• 批准号: 2181147
• 负责人: JOHN D SIMON
• 金额: $ 10.39万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-01-01 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2181147
• 关键词: band 3 protein; bioengineering /biomedical engineering; biomedical equipment development; carboxyhemoglobin; chemical association; chemical models; circular dichroism; conformation; cytochrome c; hemoglobin; hemoprotein; laser spectrometry; myoglobin; oxidation reduction reaction; photochemistry; photolysis; protein folding; protein structure function; stop flow technique; synthetic peptide; time resolved data; tryptophan; ultraviolet radiation

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.

时间分辨圆二色性（TRCD）光谱是一种新的光谱技术 我们在实验室开发的实验技术 蛋白质构象动力学研究。 该提案提出 使用TRCD和生物化学过程的持续和新的研究 互补的时间分辨激光光谱。 在资助期间，与该项目相关的两项技术发展 TRCD 技术及其在光谱学研究中的应用 将追求生物分子中的构象运动。 这是 我们的目标是开发瞬态紫外线收集技术 圆二色光谱。 我们方法的实验细节是 概述。 拟议的改进将能够收集 205 nm 至 250 nm 之间任意位置的紫外圆二色光谱 所需的时间延迟介于 30 皮秒和 10 微秒之间 光激发。 该光谱区域监测 蛋白质骨架的吸收。 紫外 CD 光谱 提供蛋白质二级结构的测量 解决方案。 关于血红素蛋白松弛的继续研究 将进行配体光解。 这期间主要的努力 下一个资助期将重点关注使用 UV CD 技术以及 各种激光光谱探头来检查 (a) 二次 蛋白质折叠和展开过程中的结构演化以及 (b) 小多肽中光诱导的螺旋-螺旋转变。 蛋白质 折叠是生物化学中的一个重要问题。 应用 时间解决的 UV CD 光谱技术为解决这个问题提供了新的解决方案 以及对与形成相关的时间尺度的令人兴奋的见解 蛋白质二级结构。 还将开展研究 开发新的光化学方法来触发蛋白质展开 和折叠工艺。