DYNAMIC NUCLEAR POLARIZATION SOLID STATE NMR SPECTROMETER FOR BIOMOLECULAR STUDIE

我们建议购买一台600 MHz的动态核极化（DNP）固态核磁共振（SSNMR）光谱仪。在过去的十年中，基于魔角旋转的固态核磁共振（MAS SSNMR）取得了快速的进展，包括许多不同种类蛋白质的光谱分配，原子分辨率结构的确定，以及其他方法无法获得的疾病相关蛋白质的应用。这些方法的更广泛的生物医学影响显然受到了SSNMR臭名昭著的低检测灵敏度的阻碍。DNP是最近建立的一种方法，通过外源性稳定自由基的照射，将检测灵敏度提高了惊人的一到两个数量级。我们展示了在制造商（Bruker BioSpin）演示现场使用新一代仪器仅在几个小时内捕获的初始光谱，之前没有使用光谱仪的经验。光谱非常令人鼓舞，证明该仪器运行稳定，足以进行多维光谱分析。此外，一些样品，代表提出的研究课题，表现出强烈的增强，并给出了高质量的光谱。一些美国国立卫生研究院资助的研究人员将收集详细的、否则无法获得的蛋白质结构信息，这些蛋白质在以下方面发挥关键作用：神经退行性变和衰老（阿尔茨海默氏症A2、突触核蛋白、tau、人类朊蛋白、肌动蛋白）、心血管疾病（钾通道和同源物）、肺部疾病（β -2-肾上腺素能受体）和传染病（HIV衣壳、金黄色金黄色球菌的多重耐药因子、结核分枝杆菌的ATP合成酶）。ESR和NMR专家将合作确保仪器的有效和创新使用。专门负责拟议仪器的工作人员将帮助传播有关方法的指导，以促进纽约结构生物学联合会10个成员机构中生物化学实验室之间的合作。)

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.