500 MHz NMR Spectrometer at the Medical College of Wisconsin

描述（由申请人提供）：本申请的总体目标是在威斯康星州医学院开发共享资源，用于使用高分辨率NMR光谱分析生物分子结构和动力学。已要求提供资金，以购买一台500兆赫核磁共振波谱仪，该波谱仪将提供给威斯康星州东南部生物医学研究界不断扩大的用户群。该工具将支持和扩展现有的NIH项目，供来自8个机构的4个主要用户和12个次要用户使用。主要项目由美国国立卫生研究院的6笔赠款资助，次要用户则得到各机构的支持。该用户组的NMR需求超过了威斯康星州医学院现有600 MHz仪器的可用时间。蛋白质动力学研究的一个关键要求是在两个不同的磁场强度下获得的弛豫数据，本申请中要求的配备冷冻探针的500 MHz仪器将提供在附近其他NMR设施中不容易获得的最佳解决方案。主要项目包括：（1）炭疽光活化素天然状态结构平衡的动力学（Dahman），（2）选择性激活炭疽杀灭酶PlyG的结构基础（Dahman），（3）真核结构基因组学中心的靶标筛选和优化（Dahman），（4）甘露糖-6-磷酸受体的结构分析（Dahms），（5）Biclycine增强SOD/ALS突变体（Kalyanaraman）的过氧化和（6）PECAM-1胞质结构域的溶液结构（纽曼）。500 MHz仪器将是MCW生物分子NMR中心重大扩建的第一个组成部分，同时计划建立一个新的结构生物学中心。因此，妇女事务部已承诺为该仪器提供250 000美元的配套资金，并为建造核磁共振设施提供500 000美元。相关性：蛋白质折叠、结构和动力学的改变是许多人类疾病的基础。使用所要求的NMR仪器的项目将回答有关导致ALS（Lou Gehrig病）或轻链淀粉样变性（AL）的遗传突变的问题，展示工程蛋白如何用于治疗溶酶体储存疾病（例如Gaucher，Fabry和Tay-Sachs），并定义选择性炭疽杀灭酶的结构。其他研究将利用磁场强度的差异来测量控制癌症和免疫反应中细胞信号的三维蛋白质结构的波动。公共卫生相关性：蛋白质折叠、结构和动力学的改变构成了许多人类疾病的基础。使用所要求的NMR仪器的项目将回答有关导致ALS（Lou Gehrig病）或轻链淀粉样变性（AL）的遗传突变的问题，展示工程蛋白如何用于治疗溶酶体储存疾病（例如Gaucher，Fabry和Tay-Sachs），并定义选择性炭疽杀灭酶的结构。其他研究将利用磁场强度的差异来测量控制癌症和免疫反应中细胞信号的三维蛋白质结构的波动。

DESCRIPTION (provided by applicant): The overall objective of this application is to develop a shared resource at the Medical College of Wisconsin for the analysis of biomolecular structure and dynamics using high-resolution NMR spectroscopy. Funds are requested for a 500 MHz NMR spectrometer, which will be made available to an expanding base of users in the biomedical research community of southeastern Wisconsin. This instrument will support and extend existing NIH projects for 4 major users and 12 minor users from 8 institutions. The major projects are funded by 6 NIH grants, and minor users are supported by various agencies. The NMR needs of this user group exceed the time available on existing 600 MHz instruments at the Medical College of Wisconsin. A key requirement for investigations of protein dynamics is relaxation data acquired at two different magnetic field strengths, and the cryoprobe-equipped 500 MHz instrument requested in this application will provide an optimal solution that is not readily available at other nearby NMR facilities. Major projects include: (1) Dynamics of the lymphotactin native state structural equilibrium (Volkman), (2) Structural basis of selective activation of the anthrax-killing enzyme PlyG (Volkman), (3) Target screening and optimization for the Center for Eukaryotic Structural Genomics (Volkman), (4) Structural analysis of mannose-6-phosphate receptors (Dahms), (5) Bicarbonate enhances peroxidation of SOD/ALS mutants (Kalyanaraman) and (6) Solution structure of the PECAM-1 cytoplasmic domain (Newman). The 500 MHz instrument will be the first component of a major expansion to the MCW Biomolecular NMR Center, in conjunction with plans for a new Center for Structural Biology. Accordingly, MCW has committed $250,000 in matching funds for the instrument and $500,000 for construction of the NMR facility. Relevance: Alterations in protein folding, structure and dynamics form the basis of many human diseases. Projects using the requested NMR instrument will answer questions about inherited mutations that cause ALS (Lou Gehrig's disease) or light-chain amyloidosis (AL), show how engineered proteins can be used to treat lysosomal storage diseases (Gaucher, Fabry, and Tay-Sachs, for example), and define the structure of a selective anthrax-killing enzyme. Other studies will exploit differences in magnetic field strength to measure fluctuations in three-dimensional protein structures that control cellular signals in cancer and the immune response. PUBLIC HEALTH RELEVANCE: Alterations in protein folding, structure and dynamics form the basis of many human diseases. Projects using the requested NMR instrument will answer questions about inherited mutations that cause ALS (Lou Gehrig's disease) or light-chain amyloidosis (AL), show how engineered proteins can be used to treat lysosomal storage diseases (Gaucher, Fabry, and Tay-Sachs, for example), and define the structure of a selective anthrax-killing enzyme. Other studies will exploit differences in magnetic field strength to measure fluctuations in three-dimensional protein structures that control cellular signals in cancer and the immune response.