Pulsed Q-band EPR Spectrometer

描述(由申请者提供)：请提供资金以完成在范德比尔特大学购买Bruker EleXsys 580 Q波段FT-EPR系统。这台设备将被五个不同的研究小组和他们的一些合作者用来对广泛的氮氧化物自旋标记蛋白质进行鹿测量，这些蛋白质已经在使用定点自旋标记进行研究。在仪器和方法学方面的最新进展将时域EPR定位为表征自旋标记蛋白质的结构和动力学的CW-EPR的重要补充。在NIH资助的构成这一应用的核心用户群的十个项目中，所要求的设备将用于：1)增加我们测量鹿蛋白质“长”(&gt；20-25 E)探针间距离的能力；2)提高小样本的信噪比，包括重组膜蛋白；3)允许对鹿数据进行多频分析；4)为评估自旋标签之间的相对取向提供方向选择性；以及5)提供可靠的CW-Q波段测量能力。在五个申请实验室中的每一个中，所要求的仪器将通过提供独特的新的测量能力来全面表征具有明显生物医学重要性的各种可溶性和膜结合蛋白的结构和动力学，从而产生直接和实质性的影响。范德比尔特和周围地区的其他调查人员也将获得所要求的仪器，因为他们正在开发涉及自旋标记的新项目。设备的公平使用将由内部监督委员会确保，日常操作将由经验丰富的EPR光谱仪处理。

DESCRIPTION (provided by applicant): Funds are requested to complete the purchase of a Bruker EleXsys 580 Q-band FT-EPR system at Vanderbilt University. This equipment will be utilized by five different established research groups and a number of their collaborators to make DEER measurements on a wide range of nitroxide spin-labeled proteins that are already being studied using site-directed spin-labeling. Recent advances in instrumentation and methodology have positioned time-domain EPR as an essential complement to cw-EPR for characterizing the structures and dynamics of spin-labeled proteins. In the ten NIH funded projects that constitute the core user group of this application, the requested equipment will be utilized to: 1) increase our capacity for measuring "long" (> 20 - 25 E) inter-probe distances in proteins by DEER; 2) increase signal to noise for small samples including reconstituted membrane proteins; 3) permit multifrequency analysis of DEER data; 4) provide orientation selectivity for assessment of relative orientations between spin labels; and 5) provide the capability to make reliable cw-Q-band measurements. In each of the five applicant laboratories, the requested instrumentation will have an immediate and substantial impact by providing unique new measurement capabilities for full characterization of the structures and dynamics of wide range of soluble and membrane bound proteins that are of clear biomedical importance. The requested instrumentation will also be made available to other investigators at Vanderbilt and in the surrounding area as they develop new projects that involve spin-labeling. Equitable utilization of the equipment will be assured by an internal oversight committee and day-to-day operation will be handled by an experienced EPR spectroscopist.