Conformational Dynamics of Ribosomal Proteins

9808427詹姆森这项研究的目的是提高对核糖体“茎”蛋白的亚基相互作用和构象动力学的理解，核糖体“茎”蛋白既是分离的，也是与其他核糖体成分相关的。待研究的具体系统包括大肠杆菌。大肠杆菌核糖体蛋白L7/L12和L10，重组为核糖体亚基和酵母真核蛋白YPO、YP2(和YP1(.半胱氨酸残基通过定点突变技术特定地放置在这些蛋白质中的适当位置，将被巯基特异的荧光探针标记。荧光方法，包括稳态和时间分辨的各向异性，然后将被用来表征这些蛋白质的不同结构域的旋转性质，这些结构域在溶液中是自由的，并被结合到核糖体中。这些后一项研究的目的是阐明二聚体/单体平衡的热力学方面，以及研究在溶液中和各种络合物中游离的二聚体之间的亚基交换。核糖体是制造新蛋白质的细胞器，因此对细胞的正常功能和生命是必不可少的。这些研究将有助于更好地了解核糖体的各个部分在执行其生物功能时是如何运动的。研究方案的培训方面也将有助于开发人力资源。

9808427 Jameson The object of this research is to improve the understanding of subunit interactions and conformational dynamics of ribosomal "stalk" proteins, both isolated and associated with other ribosomal components. The specific systems to be studied include the Escherichia. coli ribosomal proteins L7/L12 and L10, free and reconstituted into ribosomal subunits and the yeast eukaryotic proteins YPO, YP2( and YP1(. Cysteine residues, specifically placed at appropriate locations within these proteins by site-directed mutagenesis techniques, will be labeled with sulfhydryl specific fluorescence probes. Fluorescence methods, including both steady-state and time-resolved anisotropies, will then be utilized to characterize the rotational properties of different domains of these proteins, free in solution and incorporated into ribosomes. These latter studies will be aimed at elucidating thermodynamic aspects of the dimer/monomer equilibria as well as studying subunit exchange between dimers free in solution and in the various complexes. The ribosome is the cellular organelle that makes new proteins and hence is essential to the proper functioning of the cell and to life. These studies will help to provide a better understanding of how the various parts of the ribosome move as they carry out their biological function. The training aspects of the research program will also help to develop human resources.