Structure Determination by NMR

整个项目将确定生殖支原体和肺炎支原体的蛋白质结构。这些结构将增强蛋白质折叠的数据库，并将被解释为暗示蛋白质的可能功能。整个项目的这一部分将应用溶液NMR方法，使用多核多维方法确定这些生物体中小蛋白质的结构。将设计一组实验，这些实验针对具有特定线宽的蛋白质的共振分配进行优化。 将实施用于数据自动分析的方法（确定主链和侧链分配，以及NOE交叉的分配），并将其扩展以优化通量。与M.g.同源的蛋白和M.p.，但来自嗜热生物体，将在NMR工作中进行评价。由于线宽减小，在较高温度下的数据收集得到改善，提高了数据质量，从而提高了自动分析的能力。根据褶皱分类和功能确定的结构的解释将与该项目的其他成员合作完成。NMR也将被应用于评估蛋白质的结晶学，试图确定灵活的区域，和comains可能会阻碍结晶学，试图确定灵活的区域，和域可能会阻碍结晶。

The overall project will determine the structure of proteins from Mycoplasma genitalium and Mycoplasma pneumoniae. These structures will enhance the database of protein folds, and will be interpreted to suggest possible functions for the proteins. This component of the overall project will apply solution NMR methods to determine structures of small proteins from these organisms using multinuclear multidimensional methods. Sets of experiments will be designed which are optimized for assignment of resonances from proteins with specific linewidths. Methods for automated analysis of data (determination of backbone and sidechain assignments, and assignment of NOE crosspeaks) will be implemented, and extended to optimize throughput. Proteins homologous to M.g. and M.p. but from thermophilic organisms will be evaluated fro NMR work. Data collection at higher temperature is improved due to decreased linewidths, enhancing data quality and hence the ability to automate analysis. The interpretation of structures determined in terms of fold classification and function will be done in collaboration with other members of the project. NMR will also be applied to evaluation of proteins for crystallography, attempting to identify flexible regions, and comains which may hinder crystallography, attempting to identify flexible regions, and domains which may hinder crystallization.