Time-domain/ELDOR EPR Spectrometer

描述(由申请人提供)：这是一份共享仪器资助方案，用于购买现代商业CW和FT-EPR Bruker EleXsys X波段光谱仪，供北卡罗来纳州研究三角地区大学的六名资深首席研究员和一名初任研究员组成的核心小组使用。目前，三角地区还没有这样的仪器。正在进行的研究项目涉及现代时域EPR方法的应用，包括脉冲饱和恢复和HYSCORE在广泛的生物医学问题和系统中的应用，从膜蛋白生物芯片的混合纳米级组装到脂转移蛋白，tRNA的自组装和纳米粒子模板病毒粒子辅助的药物输送，以及酶的催化部位的结构。目前，所有这些项目要么是使用已有至少20年历史的现有连续波X波段EPR仪器进行的，要么是与拥有Bruker EleXsys X波段光谱仪的其他组织合作进行的。随着这次拟议的重大升级，所有七个核心研究项目都将受益于X波段时间域EPR方法的仪器和方法学的最新发展。具体地说，我们将能够1)获得大生物分子组装和部分折叠结构的长程(高达60-70埃)距离约束；2)在顺磁松驰剂存在和不存在的情况下，直接测量特定标记的蛋白质、RNA和磷脂的电子松弛速率，以获得有价值的结构信息；和3)获得关于底物与催化酶结合时顺磁性金属离子配位的独特信息(HYSCORE实验)。PUBLIC健康相关性：该项目与公共健康的相关性将主要是为NIH资助的研究人员以及来自NCSU、北卡罗来纳大学和研究三角地区的其他研究人员提供先进的FT-EPR光谱仪，因为这些研究人员正在开发涉及蛋白质和RNA结构功能研究的新项目。新的FT-EPR光谱仪还将加强对新设立的NCSU RNA生物学/化学-生物学接口培训项目的研究生的培训，该项目涉及NCSU的十个系。

DESCRIPTION (provided by applicant): This is a Shared Instrumentation Grant proposal for the acquisition of a modern commercial CW and FT-EPR Bruker EleXsys X-band spectrometer for use by a core group of six established Principal Investigators and one beginning investigator from Universities in the Research Triangle area of North Carolina. Currently, there is no such instrument in the Triangle area. The ongoing research projects involve applications of modern time-domain EPR methods including pulsed saturation recovery and HYSCORE to a wide range of biomedical problems and systems from hybrid nanoscale assemblies for membrane protein biochips to lipid transfer proteins, self-assembly of tRNA and drug-delivery assisted by nanoparticle-templated virions, and the structures of the enzymes' catalytic sites. Currently, all these projects are either pursued with available continuous wave X-band EPR instruments that are at least 20 years old or in collaborating with other groups that have Bruker EleXsys X-band spectrometer available. With this proposed major upgrade all seven core research projects will benefit from recent developments in instrumentation and methodology of time-domain EPR methods at X-band. Specifically, we will be able to 1) obtain long-range (to up to 60-70 angstrom) distance constrains for large biomolecular assemblies and partially folded structures; 2) directly measure electronic relaxation rates of specifically labeled proteins, RNAs, and phospholipids in the presence and absence of paramagnetic relaxation agents in order to gain valuable structural information; and 3) obtain unique information on paramagnetic metal ion coordination upon substrate binding to catalytic enzymes (HYSCORE experiment).PUBLIC HEALTH RELEVANCE: The relevance of this project to public health will be in providing primarily NIH-funded investigators as well as other investigators from NCSU, UNC, and the Research Triangle area with an advanced FT- EPR spectrometer as these investigators develop new projects involving structure-function studies of proteins and RNAs. The new FT-EPR spectrometer will also enhance training of graduate students from the newly established NCSU RNA Biology/Chemistry -Biology Interface Training Program that involves ten NCSU Departments.