DYNAMIC NUCLEAR POLARIZATION SOLID STATE NMR SPECTROMETER FOR BIOMOLECULAR STUDIE

用于生物分子研究的动态核偏振固态核磁共振波谱仪

• 批准号: 7839443
• 负责人: ANN E MCDERMOTT
• 金额: $ 230.93万
• 依托单位: NEW YORK STRUCTURAL BIOLOGY CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-20 至 2013-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7839443
• 关键词: Actins; Aging; Alzheimer' s Disease; American; Biochemistry; Capsid; Cardiovascular Diseases; Collaborations; Communicable Diseases; Detection; Disease; Ensure; Exhibits; Free Radicals; Funding; Generations; HIV; Homologous Gene; Hour; Human; Institution; Instruction; Laboratories; Lung diseases; Magic; Manufacturer Name; Methods; Multi-Drug Resistance; Mycobacterium tuberculosis; Nerve Degeneration; New York; Nuclear; Occupations; Play; Potassium Channel; Prions; Proteins; Research; Research Personnel; Resolution; Role; Sampling; Site; Spectrum Analysis; Staphylococcus aureus; Structure; base; beta-2 Adrenergic Receptors; experience; improved; innovation; instrument; instrumentation; irradiation; member; public health relevance; resistance factors; solid state; solid state nuclear magnetic resonance; structural biology; synuclein; tau Proteins

DESCRIPTION (provided by applicant): DNP SSNMR Spectrometer for Biomolecular Studies We propose the purchase of a Dynamic Nuclear Polarization (DNP) Solid State NMR (SSNMR) spectrometer at 600 MHz. Over the past decade Magic Angle Spinning based Solid State NMR (MAS SSNMR) has made rapid progress, including spectral assignments of many different kinds of proteins, determination of atomic-resolution structures, and applications to disease related proteins that are inaccessible by other methods. Broader biomedical impact of these methods is clearly hampered by the notoriously poor detection sensitivity of SSNMR. DNP is a recently established method to improve detection sensitivity by an astonishing one to two orders of magnitude, via irradiation of exogenous stable free radicals. We present initial spectra, captured in only a few hours on this new-generation instrument at the manufacturer's (Bruker BioSpin) demonstration site, with no previous experience with the spectrometer. The spectra are very encouraging, validating that the instrumentation operates stably enough for multidimensional spectroscopy. Moreover, several samples, representative of the proposed research topics, exhibit strong enhancements, and give high quality spectra. Several NIH-funded investigators will gather detailed, otherwise unavailable, structural information on proteins that play key roles in: neurodegeneration and aging (Alzheimer's A2, synuclein, tau, human prion protein, actin), cardiovascular disease (potassium channels and homologs), pulmonary disease (beta-2-adrenergic receptor) and infectious diseases (HIV capsid, multiple drug resistance factor from S. aureus, ATP synthase from M. tuberculosis). ESR and NMR experts will collaborate to ensure effective and innovative use of the instrument. Staff devoted to the proposed instrument will help disseminate instruction on the methods, to facilitate collaboration among biochemistry laboratories in the 10 member institution of the New York Structural Biology Consortium. ) PUBLIC HEALTH RELEVANCE: We propose the purchase of a new-generation Dynamic Nuclear Polarization Solid State NMR spectrometer at 600 MHz, which will provide detailed, otherwise unavailable, structural information on proteins that play key roles in neurodegeneration and aging, cardiovascular disease, pulmonary disease and infectious diseases. On funding, the purchase would protect American jobs, both in the American corporate supplier (Bruker BioSpin in Billerica, MA) and in the host laboratory, a consortium of 10 member institutions of the New York Structural Biology Consortium.

描述（由申请人提供）：用于生物分子研究的 DNP SSNMR 波谱仪 我们建议购买 600 MHz 的动态核极化 (DNP) 固态 NMR (SSNMR) 波谱仪。在过去的十年中，基于魔角旋转的固态核磁共振 (MAS SSNMR) 取得了快速进展，包括许多不同种类蛋白质的光谱分配、原子分辨率结构的测定以及对其他方法无法获得的疾病相关蛋白质的应用。众所周知，SSNMR 的检测灵敏度较差，这显然阻碍了这些方法更广泛的生物医学影响。 DNP 是最近建立的一种方法，通过外源稳定自由基的照射，将检测灵敏度提高了惊人的一到两个数量级。我们展示了初始光谱，这些光谱是在制造商 (Bruker BioSpin) 演示现场的新一代仪器上仅用几个小时捕获的，之前没有使用光谱仪的经验。光谱非常令人鼓舞，验证了仪器对于多维光谱学来说足够稳定地运行。此外，代表所提出的研究主题的几个样品表现出强大的增强功能，并提供高质量的光谱。 NIH 资助的几位研究人员将收集有关蛋白质的详细的、否则无法获得的结构信息，这些蛋白质在以下方面发挥关键作用：神经退行性疾病和衰老（阿尔茨海默氏症 A2、突触核蛋白、tau、人朊病毒蛋白、肌动蛋白）、心血管疾病（钾通道和同源物）、肺部疾病（β2 肾上腺素能受体）和传染病（HIV 衣壳、 来自金黄色葡萄球菌的多重耐药因子、来自结核分枝杆菌的 ATP 合酶）。 ESR 和 NMR 专家将合作确保该仪器的有效和创新使用。致力于拟议仪器的工作人员将帮助传播有关方法的说明，以促进纽约结构生物学联盟 10 个成员机构的生物化学实验室之间的合作。 ） 公共健康相关性：我们建议购买新一代动态核极化固态核磁共振波谱仪，频率为 600 MHz，它将提供有关在神经退行性疾病和衰老、心血管疾病、肺部疾病和传染病中发挥关键作用的蛋白质的详细的、否则无法获得的结构信息。在资金方面，此次收购将保护美国的就业机会，包括美国企业供应商（位于马萨诸塞州比尔里卡的布鲁克 BioSpin）和主办实验室（由纽约结构生物学联盟的 10 家成员机构组成的联盟）的就业机会。