表征天然丰度酵母细胞色素c多构象的液体14N NMR方法研究

项目名称： 表征天然丰度酵母细胞色素c多构象的液体14N NMR方法研究

项目编号： No.21505153

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 数理科学和化学

项目作者： 孙鹏

作者单位： 中国科学院武汉物理与数学研究所

项目金额： 21万元

中文摘要： 蛋白质功能相关的多构象现象正成为新的研究热点。核磁共振波谱（NMR）是一种非常有效的获得生物大分子原子水平结构及动力学信息的技术.然而，对于同时存在多种构象的蛋白，由于很难获得与某一特定功能相关的单一构象，通常需要外源标记技术以区分不同的构象，这些方法往往操作复杂、费用昂贵、而且可能影响目标蛋白的固有性质。因此，发展新的内源探针来表征多构象蛋白具有非常重要的意义。细胞色素c是一种非常典型的存在多种构象且行使多种生物学功能的蛋白质，同时在呼吸作用和细胞凋亡中起重要作用，研究细胞色素c的多构象现象对于人们理解细胞色素c如何发挥其功能有重要的帮助。本项目中，申请人将发展基于液体14N NMR的蛋白质动力学表征方法，并以此为探针研究天然丰度酿酒酵母iso-1细胞色素c功能相关的构象转变。项目的顺利实施会帮助人们理解细胞色素c结构与功能的关系，也将为天然提取蛋白质的动力学研究提供新的技术方法和路线。

中文关键词： 细胞色素c；14N核磁共振波谱；多重构象；天然丰度；蛋白质动力学

英文摘要： Functionally related conformational heterogeneity of proteins is becoming a new research hotspot. NMR is a powerful probe for the study of protein structure and dynamics with atomic resolution. However, it is hard to obtain a single conformer related to a specific function for proteins with conformational heterogeneity. Exogenously selective labeling methods are usually employed to differentiate different conformers, which may be complex, costly and lead to results deviated from original facts. Therefore, developing new endogenous probe to study functionally related conformational heterogeneity of proteins is meaningful. Cytochrome c is a representative protein, which possesses several conformers exhibiting important biological functions in respiration and apoptosis. Exploring functionally related conformational heterogeneity of cytochrome c is helpful to understand the relevant mechanism accurately. In this project, we will develop liquid 14N NMR based methods to study functionally related conformational heterogeneity of yeast iso-1-cytochrome c at natural abundance. The implement of this project will be helpful for understanding the relation between structures and functions of yeast iso-1-cytochrome c and meaningful for making liquid 14N NMR to be a new endogenous probe for protein dynamics at natural abundance.

英文关键词： cytochrome c;14N NMR;conformational heterogeneity;natural abundance;protein dynamics