CAREER: Elucidation of the Conformation and Dynamics of Membrane Proteins by Solid-State NMR

Hing，MeiMCB-0093398本研究的主要目的是使用先进的固态NMR光谱来阐明膜蛋白的结构和动力学。膜蛋白调节细胞转运和细胞信号传导，因此它们对细胞存活是必不可少的.了解它们的三维结构对于理解它们的功能至关重要。固体核磁共振技术是研究膜蛋白在脂双层中分子结构的一种很有前途的方法，在这个CAREER项目中，我们将利用适合广泛同位素标记蛋白质的核磁共振技术研究22 kDa膜结合蛋白--colicinIa通道结构域的结构和动力学。大肠杆菌素中的a-螺旋和b-折叠片段将被鉴定，具体的结构约束将使用化学位移各向异性、相关偶极耦合和1H-1H核间距来测量。蛋白质插入脂质双层的深度将通过1H自旋扩散技术测定。大肠杆菌素主链和侧链的动力学将使用各向异性耦合，线形和弛豫时间常数来表征。这些研究将有助于深入了解大肠杆菌素Ia和其他蛋白质的构象变化机制，这些蛋白质通过重折叠来实现其功能。与这些研究目标相关的是本科化学课程现代化的教育举措。将设计普通化学的蛋白质结构模块，将研究与教学结合起来，并为本科物理化学课程创建网页。将通过在实验室招聘女性暑期实习生来鼓励妇女参与化学。

Hing, MeiMCB-0093398The broad objective of this research is to elucidate membrane protein structure and dynamicsusing advanced solid-state NMR spectroscopy. Membrane proteins regulate cellular transport andcell signaling, thus they are essential for cell survival. Knowledge of their three-dimensionalstructures is crucial for understanding their functions. Solid-state NMR is a promising method forstudying the molecular structure of membrane proteins in their native environment of lipid bilayers.In this CAREER project, the structure and dynamics of a 22 kDa membrane-bound protein, colicinIa channel domain, will be investigated using NMR techniques suitable for extensively isotopicallylabeled proteins. a-helical and b-sheet segments in colicin will be identified and specific structuralconstraints will be measured using chemical shift anisotropies, correlated dipolar couplings, and 1H-1H internuclear distances. The depth of insertion of the protein into the lipid bilayer will bedetermined by 1H spin diffusion techniques. The dynamics of the colicin backbone and sidechainwill be characterized using anisotropic couplings, lineshapes, and relaxation time constants. Thesestudies should provide insight into the mechanism of conformational changes of colicin Ia and other proteins that refold to accomplish their functions. Related to these research objectives areeducational initiatives to modernize the undergraduate chemistry curriculum. A protein structure module for general chemistry will be designed to integrate research with teaching, and create webpages for undergraduate physical chemistry courses. The participation of women in chemistry will be encouraged by recruiting women summer interns into the laboratory.