类细胞环境下蛋白质结构与功能的NMR 研究
NMR Study of Protein Structure and Function in Cell-Like Environment

徐国华李从刚 ^* 刘买利 ^*

文摘	细胞环境是大多数蛋白质行使其功能的天然环境。在细胞环境中,大分子拥挤、限域、弱相互作用等普遍存在,已经有越来越多的研究表明这种复杂细胞环境会影响蛋白质的结构与功能。因此,在越接近天然的环境中研究蛋白质,越有可能揭示蛋白质功能的本质。蛋白质结构与功能的原位研究已是目前蛋白质科学中的前沿领域。核磁共振波谱技术是研究复杂环境中蛋白质结构与功能最具潜力的手段,它能提供原子分辨的蛋白质结构,动态与相互作用信息。本文总结了我们近几年利用核磁共振技术研究大分子拥挤、限域、以及原核E. coli细胞、非洲爪蟾卵母细胞中蛋白质结构与功能等方面取得的重要进展。
其他语种文摘	Most proteins function in the cell where macromolecular crowding,confinement and quinary interaction are ubiquitous. More and more researches suggest that the complex cellular environment affects proteins structure and function. Therefore,for protein studies,the closer to native cellular environment,the more likely molecular mechanisms of proteins function could be revealed accurately. In-cell study of protein structure and function has been a frontier topic in protein science. Nuclear magnetic resonance ( NMR) spectroscopy is the most promising technique for protein structural and functional assay at atomic level in complex environments. Here, we summarize our recent progress in protein structural and functional studies in macromolecular crowding,confinement,prokaryotic and eukaryotic cells by NMR spectroscopy. Two model proteins,an intrinsic disordered protein α-synuclein and a multi-domain protein calmodulin,are employed to show how macromolecular crowding and confinement affect protein structure and function,respectively. Then incell NMR methods, including labeling strategy, cytoplasmic viscosity measurement, quinary interaction quantification are developed to obtain high-quality NMR spectra for facilitating protein structural and functional studies in living cells. Ca~(2+) -induced calmodulin conformational transitions and GB1 protein structural determination in living Xenopus laevis oocytes,are shown here as typical applications of in-cell NMR. Finally, the conclusion and perspective of environmental effects on protein structure and function are presented.
来源	化学进展 ,2017,29(1):75-82 【核心库】
DOI	10.7536/pc161225
关键词	核磁共振波谱 ; 大分子拥挤 ; 限域 ; 五级相互作用 ; 蛋白质的结构和功能
地址	中国科学院武汉物理与数学研究所, 中国科学院生物磁共振分析重点实验室;;波谱与原子分子物理国家重点实验室;;生命分析化学协同创新中心, 武汉, 430071
语种	中文
文献类型	研究性论文
ISSN	1005-281X
学科	化学;细胞生物学;生物化学
基金	国家973计划 ; 国家自然科学基金项目
文献收藏号	CSCD:5951249