Integrated state-of-the-art LC-MS/NMR system for the University of Wisconsin

描述（由申请人提供）：申请资金用于购买一套集成的液相色谱、质谱仪和核磁共振（LC-MS/NMR）系统。该系统将被安置在位于威斯康星大学麦迪逊分校生物化学系的麦迪逊国家磁共振设施（NMRFAM）中。该系统将安装在一个严格控制温度和湿度的房间里，所有需要的设施都已经到位。该系统将用于支持正在进行的nih支持的四个广泛领域的研究项目：(1)代谢物混合物分析（代谢组学）。(2)药物受体（配体-蛋白）相互作用的筛选。(3)通过核磁共振筛选蛋白质和核酸样品，在各种条件下进行适当的折叠，并通过质谱进行质量控制（正确的识别、标记和纯度）。(4)阐明少量天然产物的结构。LC-MS/NMR将具有该校园或目前美国任何其他学术设施无法比拟的独特功能。该系统将具有自动化，多样品操作与样品温度控制。它将有能力获取ESI/APCI Q-TOF质谱或质谱/质谱数据的等分样品（未分馏或LC部分），也可以通过核磁共振光谱进行研究。核磁共振光谱仪将由一块600兆赫的屏蔽磁铁、一个多核控制台和一个低温平台组成。用户可以选择两种三共振（1H或13C观察，13C或15N去耦，2H锁定）低温探头：对于足够的样品，5mm探头将提供最高灵敏度(300 ？L样管)；对于样品有限的情况，1.7 mm探头将提供卓越的性能(30 ？L试管样品)。这种核磁共振系统将使蛋白质和核酸的经济筛选和天然产物的研究只有在小批量获得。该系统将有一个固相萃取附件，能够浓缩样品高达10倍。液体处理系统将样品转移到质谱系统或核磁共振管。恒温换样器将处理管中核磁共振样品的自动筛选。该系统将有一个软件系统，用于自动识别和量化代谢物和相关的生物合成途径。NMRFAM工作人员将维护仪器，协助进行研究，并培训希望获得如何安全正确使用系统的实践经验的用户。收取用户费用以收回营运开支。日程安排将由现有的网络系统进行，监督将由现有的地方和外部咨询委员会提供。公共卫生相关性：该仪器支持的研究预计将有益于公共卫生，因为它旨在调查衰老和各种疾病状态的代谢标志物，并确定候选新药和抗生素的天然产物的结构。该仪器将能够更全面地研究与人类健康有关的蛋白质和核酸的结构，以及它们如何与药物和调节其活性的天然分子相互作用。

DESCRIPTION (provided by applicant): Funds are requested for the purchase of an integrated liquid-chromatography, mass spectrometer, and nuclear magnetic resonance (LC-MS/NMR) system. This system will be housed in the National Magnetic Resonance Facility at Madison (NMRFAM) located in the Biochemistry Department at the University of Wisconsin-Madison. The system will be installed in a room with tightly controlled temperature and humidity and with all needed utilities already in place. The system will be used to support ongoing, NIH-supported, research projects in four broad areas: (1) Analysis of metabolite mixtures (metabolomics). (2) Screening of drug-receptor (ligand- protein) interactions. (3) Screening of protein and nucleic acid samples by NMR for proper folding under various conditions and by MS for quality control (correct identity, labeling, and purity). (4) Elucidation of structures of natural products available only in small quantities. The LC-MS/NMR will have unique capabilities unmatched on this campus or at currently any other academic facility in the US. The system will have automated, multiple sample operation with temperature control of samples. It will have the ability to acquire ESI/APCI Q-TOF MS or MS/MS data on aliquots of samples (unfractionated or LC fraction) that can be investigated also by NMR spectroscopy. The NMR spectrometer will consist of a 600 MHz shielded magnet, a multinuclear console, and a cryogenic platform. Users will have the choice of two triple-resonance (1H or 13C observe, 13C or 15N decoupling, 2H lock) cryogenic probes: for adequate samples, a 5 mm probe will provide the highest sensitivity (300 ?L samples in tubes); for sample-limited situations, a 1.7 mm probe will provide superior performance (30 ?L samples in tubes). This NMR system will enable the economical screening of proteins and nucleic acids and investigations of natural products obtainable only in small quantities. The system will have a solid-phase extraction accessory capable of concentrating samples by up to 10-fold. A liquids handling system will transfer samples to the MS system or NMR tubes. A thermostatted sample changer will handle the automated screening of NMR samples in tubes. The system will have a software system for automated identification and quantification of metabolites and related biosynthetic pathways. NMRFAM staff members will maintain the instrumentation, assist in performing studies, and train users who desire hands-on experience how to use the system safely and properly. User fees will be levied to recover operating expenses. Scheduling will be by an existing web-based system, and oversight will be provided by existing local and external advisory committees. PUBLIC HEALTH RELEVANCE: The research supported by this instrumentation is expected to benefit public health because it aims to investigate metabolic markers of aging and various disease states and to determine the structures of natural products that are candidate new drugs and antibiotics. The instrumentation will enable more complete investigations of the structures of proteins and nucleic acids related to human health and how they interact with drugs and naturally occurring molecules that modulate their activity.