500 MHz NMR Spectrometer at the Medical College of Wisconsin

威斯康星医学院 500 MHz 核磁共振波谱仪

• 批准号: 7595683
• 负责人: Brian F Volkman
• 金额: $ 50万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7595683
• 关键词: Amyloidosis; Amyotrophic Lateral Sclerosis; Anthrax disease; Bicarbonates; Biomedical Research; Biomolecular Nuclear Magnetic Resonance; CD31 Antigens; Commit; Communities; Cytoplasmic Tail; Data; Enzymes; Equilibrium; Funding; Grant; IGF Type 2 Receptor; Immune response; Inherited; Institution; Investigation; Lysosomal Storage Diseases; Malignant Neoplasms; Measures; Minor; Mutation; NMR Spectroscopy; Protein Dynamics; Protein Engineering; Relaxation; Resolution; Resource Sharing; Screening procedure; Signal Transduction; Solutions; Structure; Time; United States National Institutes of Health; Wisconsin; base; human disease; instrument; killings; magnetic field; medical schools; mutant; peroxidation; primary amyloidosis of light chain type; protein folding; protein structure; public health relevance; structural biology; structural genomics

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.

描述（由申请人提供）：本申请的总体目标是在威斯康星医学院开发共享资源，用于使用高分辨率核磁共振波谱分析生物分子结构和动力学。需要资金购买一台 500 MHz 核磁共振波谱仪，该仪器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。该工具将为来自 8 个机构的 4 个主要用户和 12 个次要用户支持和扩展现有的 NIH 项目。主要项目由 6 项 NIH 拨款资助，次要用户则得到各个机构的支持。该用户组的 NMR 需求超出了威斯康星医学院现有 600 MHz 仪器的可用时间。研究蛋白质动力学的一个关键要求是在两种不同磁场强度下获取弛豫数据，并且本应用中要求的配备冷冻探针的 500 MHz 仪器将提供最佳解决方案，而附近其他 NMR 设施无法轻易获得。主要项目包括：（1）淋巴趋化素天然状态结构平衡动力学（Volkman），（2）选择性激活炭疽杀灭酶PlyG的结构基础（Volkman），（3）真核中心靶标筛选与优化结构基因组学 (Volkman)，(4) 6-磷酸甘露糖受体的结构分析 (Dahms)，(5) 碳酸氢盐增强过氧化作用SOD/ALS 突变体 (Kalyanaraman) 和 (6) PECAM-1 胞质结构域的溶液结构 (Newman)。 500 MHz 仪器将成为 MCW 生物分子 NMR 中心重大扩建的第一个组成部分，同时还计划新建一个结构生物学中心。因此，MCW 承诺为该仪器提供 250,000 美元的配套资金，并为 NMR 设施的建设提供 500,000 美元。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（卢伽雷氏病）或轻链淀粉样变性 (AL) 的遗传突变的问题，展示如何使用工程蛋白来治疗溶酶体贮积病（Gaucher、Fabry 和 Tay-Sachs） ，例如），并定义了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。公共健康相关性：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（卢伽雷氏病）或轻链淀粉样变性 (AL) 的遗传突变的问题，展示如何使用工程蛋白来治疗溶酶体贮积病（Gaucher、Fabry 和 Tay-Sachs） ，例如），并定义了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。