600MHZ NMR SPECTROMETER

这是一份500,000美元的修订申请，用于购买一台四通道瓦里安600 MHz Inova核磁共振光谱仪，带有39通道Resonance Research Inc.垫片和Nalorac 5 mm四极共振(1H/13C/13N/31P(探头))。较高的场强将提供研究大于或等于20 kDa的蛋白质和研究RNA：蛋白质复合体(大于或等于18 kDa)所需的分辨率。包括氚在内的四个通道将使人们有可能对预氚化蛋白质进行四重共振实验，并在RNA应用中使用1H/13C/15N/31P核。此外，现有的500 MHz仪器需要19F/1H冷冻探头，以测量由19F-酪氨酸或19F-色氨酸仪器观察到的实时蛋白质折叠，以测量由19F-酪氨酸或19F-色氨酸观察到的实时蛋白质折叠。购买一台600 MHz的仪器将提供更大系统所需的灵敏度和分辨率，此外还提供松弛实验所需的另一个磁场强度，并只需提供急需的机器时间。冷冻探头可以在蛋白质(展开)的过程中快速收集核磁共振光谱。研究项目和培训。来自医学院两个系的四个实验室小组组成了一个核磁共振研究小组，其合作范围从医学院消化疾病研究核心中心的结构生物学核心，到丹福斯行星生物学研究所(孟山都/华盛顿大学)。每个小组使用核磁共振来确定蛋白质结构并观察配体的结合，包括RNA(凯瑟琳·霍尔)、脂类(David Cistola)、维甲酸(Ellen Li)和小酶辅助因子(Carl Frieden)，蛋白质和RNA动力学也是这些研究的中心，使用15N(蛋白质)或13C(RNA)松弛来表征系统的运动。

This is a revised request for $500,000 towards the purchase of a four channel Varian 600 MHz Inova NMR spectrometer, with a 39 channel Resonance Research Inc shim set and a Nalorac 5mm quadrupole resonance (1H/13C/13N/31P( probe. The higher field strength will give the resolution required for study of proteins greater than or equal to 20 kDa and for RNA: protein complexes (greater than or equal to 18 kDa). Four channels, including deuterium, will make it possible to perform quadruple resonance experiments with predeuterated proteins, as well as use 1H/13C/15N/31P nuclei in RNA applications. In addition, a 19F/1H cryoprobe is requested for the existing 500 MHz instrument, to measure real-time protein folding observed by 19 F-tyrosine or 19F-tryptophan instrument, to measure real-time protein folding observed by 19F- tyrosine or 19F-tryptophan. Acquisition of a 600 MHz instrument will give the sensitivity and resolution required for the larger systems, in addition to providing another field strength necessary for relaxation experiments, and simply giving desperately needed machine time. The cryoprobe allows rapid collection of NMR spectra during the course of a protein (unfolding). Research Projects and training. Four laboratory groups from two departments in the Medical School comprise an NMR research group with collaborations that extend from the Structural Biology Core of the Digestive Diseases Research Core Center at the Medical School, to the Danforth Planet Biology Institute (Monsanto/Washington University). Each group uses NMR to determine protein structure and to observe binding of ligands, including RNA (Kathleen Hall), lipids (David Cistola), and retinoic acid (Ellen Li) and small enzyme co-factors (Carl Frieden), Protein and RNA dynamics are also central to these investigations, using either 15N (proteins) or 13C (RNA) relaxation to characterize the motions of a system.