NOVEL 199HG NMR METHODS FOR MONITORING PROTEIN FOLDING

描述：（改编自申请人的摘要）蛋白质折叠的异常与越来越多的人类疾病有关。这些异常的起源在原子水平上的表征受到阻碍的困难，在折叠过程中监测特定的相互作用。核磁共振（NMR）光谱是研究蛋白质折叠的少数几种技术之一，可以提供原子水平的信息。由于蛋白质中富含NMR活性质子，因此所产生的基于质子的NMR光谱很少完全可分辨。需要辅因子折叠的蛋白质类提供了通过监测辅因子的NMR信号来研究折叠过程的机会。建议的工作将集中在使用199 Hg作为探针的金属蛋白质折叠。将开发二维{1H-199 Hg} NMR方法以突出折叠过程中的生物分子相互作用。这些研究还将为调查汞和某些其他金属环境暴露的病理学提供一种新的工具。X射线晶体学方法将被用来确定不同的图书馆的Hg（II）配位化合物的结构，使用蛋白质金属结合位点的合成配体模型。这个图书馆将扩展我们的知识，生物相关的结构光谱关系199 Hg化学位移和耦合常数之间的199 Hg和1H在溶液和固体状态。将制备具有良好核磁共振性能的其他过渡金属的同构配合物，以启动对其作为金属生物探针的相对优点的系统比较。最后，汞（II）配位化学的各种潜在的金属配位环二肽将使用X-射线晶体学和核磁共振检查作为前奏，这些新技术的应用，金属蛋白。

DESCRIPTION: (Adapted from the applicant's abstract) Abnormalities in protein folding are associated with a growing list of human diseases. Characterization of the origin of these abnormalities at the atomic level is hindered by difficulties in monitoring specific interactions during the folding process. Nuclear magnetic resonance (NMR) spectroscopy is one of the few techniques employed for the study of protein folding that can provide information at the atomic level. Because proteins are very rich in NMR-active protons, the resulting proton-based NMR spectra are rarely fully resolvable. The class of proteins that require cofactors for folding provide an opportunity to study the folding process by monitoring the NMR signal of the cofactor. The proposed work will focus on the use of 199Hg as a probe of metalloprotein folding. Two-dimensional {1H-199Hg} NMR methods will be developed to highlight the biomolecular interactions during the folding process. These studies will also provide a new tool for investigating the pathology of environmental exposure to mercury and certain other metals. X-ray crystallographic methods will be employed to determine the structures of a diverse library of Hg(II) coordination compounds using synthetic ligand models of protein metal-binding sites. This library will extend our knowledge of biologically relevant structure-spectroscopy relationships for 199Hg chemical shifts and coupling constants between 199Hg and 1H in solution and the solid state. Isostructural complexes of other transition metals with favorable NMR properties will be prepared to initiate a systematic comparison of their relative merits as metallobioprobes. Finally, the Hg(II) coordination chemistry of a variety of potentially metal-coordinating cyclic dipeptides will be examined using X-ray crystallography and NMR as a prelude to the application of these new techniques to metalloproteins.