CAREER: Residue Specific Probes of the Cytochrome c Folding Mechanism

职业：细胞色素 c 折叠机制的残基特异性探针

• 批准号: 0346967
• 负责人: Floyd Romesberg
• 金额: $ 79.78万
• 依托单位: The Scripps Research Institute
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-15 至 2009-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0346967&HistoricalAwards=false
• 关键词: CAREER; Residue; Specific; Probes; Cytochrome

The objective of this CAREER project is to develop a detailed model for cytochrome c folding into its native three dimensional structure. The PI's approach is based on developing a probe that is sensitive to the folding process, and may therefore be used for its characterization. First, residue-specific detail of the structural changes that accompany cytochrome c folding will be generated, both under equilibrium and kinetic folding conditions by replacing cytochrome c C-H bonds with C-D bonds. The C-D bonds are non-perturbative and sensitive to their local environment. Most importantly, the C-D bond absorbs infrared light in a region of the spectrum that is completely free of other protein or denaturant absorptions, and may thus be easily observed and characterized. To characterize the folding of cytochrome c, the C-D spectra will be characterized as the protein folds, both under equilibrium conditions and under kinetic conditions. Specific attention will be given to how different parts of the protein interact with each other during the folding process, but also to how the protein interacts with a bound iron atom which is known to be important not only for function, but for folding as well. Computer simulation studies will be pursued to facilitate the understanding of the experimental observations. The combined theoretical and experimental approach should define the cytochrome c folding process with unprecedented clarity. The techniques developed to characterize cytochrome c folding will be applicable to other proteins, and thus, these studies will have an impact on a broad range of other areas of biophysics. The educational activities in this CAREER project include training students at the interface of Chemistry and biology and expansion of internet based learning tools. This project is jointly funded by the Molecular Biophysics Program in the Division of Molecular and Cellular Biosciences and the Experimental Physical chemistry Program in the Chemistry Division.

该 CAREER 项目的目标是开发细胞色素 c 折叠为其天然三维结构的详细模型。 PI 的方法基于开发对折叠过程敏感的探针，因此可用于其表征。 首先，在平衡和动力学折叠条件下，通过用 C-D 键替换细胞色素 c C-H 键，将产生伴随细胞色素 c 折叠的结构变化的残基特异性细节。 C-D 债券不受扰动且对其当地环境敏感。 最重要的是，C-D 键吸收光谱区域中的红外光，该区域完全没有其他蛋白质或变性剂吸收，因此可以轻松观察和表征。 为了表征细胞色素 c 的折叠，C-D 光谱将表征为蛋白质折叠，无论是在平衡条件下还是在动力学条件下。 我们将特别关注蛋白质的不同部分在折叠过程中如何相互作用，以及蛋白质如何与结合的铁原子相互作用，已知这不仅对于功能很重要，而且对于折叠也很重要。 将进行计算机模拟研究，以促进对实验观察结果的理解。 理论和实验相结合的方法应该以前所未有的清晰度定义细胞色素 c 折叠过程。 为表征细胞色素 c 折叠而开发的技术将适用于其他蛋白质，因此，这些研究将对生物物理学的其他广泛领域产生影响。该职业项目的教育活动包括对学生进行化学和生物学的培训以及基于互联网的学习工具的扩展。该项目由分子与细胞生物科学部分子生物物理项目和化学部实验物理化学项目联合资助。