ELECTRON PARAMAGNETIC RESONANCE SPECTROMETER

布鲁克ER300-10/12型EPR光谱仪及其随身配件 X和Q波段输出EPR和饱和转移(ST-EPR)测量 将购买生物样本上的。这件乐器是最理想的 适合于结构的自旋标记和自旋探针研究， 可溶性和膜结合蛋白的动力学和相互作用， 上述实验室的主要研究重点课题 用户组。所需设备的采购将具有 对七个项目中的每一个都有直接和重大的影响 列在本建议书中。这些措施包括：1)更多地获得EPR 用于进行常规X波段测量的仪器 生物样品，2)获取硬件以获得更高的 频率{O-band)测量可实现更严格的分析 来自许多研究和检测的自旋标记数据 蛋白质结合的Mn+：在其他方面，3)产生高信号的能力 在色散模式下的去噪声ST-EPR测量，4)访问 用于优化的选定塞曼场调制频率 ST-EPR测量对旋转角度变化的敏感性 自旋标记蛋白质的扩散和数据分析 ST-EPR实验中的多运动种群，以及5)支持 用于光谱仪及其附件的维护和维修。 电子顺磁共振波谱目前被每一位申请者用于 围绕结构-功能这一中心主题进行的研究 蛋白质之间的关系。这个团体有一个既定的记录 新型自旋标记试剂的设计和化学合成 蛋白质的定点修饰和开发 井下资料分析的理论方法 定义的系统。EPR工作得到了各种其他工作的补充 生物物理方法，包括核磁共振(核磁共振)， X射线结晶学/分子模拟、荧光 光谱学和分子生物学/蛋白质工程。这个 申请者有共享仪器的既定记录 在他们的研究项目中进行积极的合作。

A Bruker ER300-10/12 EPR spectrometer and accessories for carrying out X- and Q-band EPR and saturation transfer (ST-EPR) measurements on biological samples will be purchased. This instrument is ideally suited for spin label and spin probe studies of the structures, dynamics and interactions of soluble and membrane bound proteins, topics of major research emphasis in the laboratories of the above user group. Acquisition of the requested equipment will have an immediate and significant impact on each of the seven projects listed in this proposal. These include: 1) increased access to EPR instrumentation for making routine X-band measurements on biological samples, 2) access to hardware for making higher frequency {O-band) measurements enabling more rigorous analysis of spin labeling data from a number of the studies and detection of protein bound Mn+: in others, 3) capability to make high signal- to-noise ST-EPR measurements in the dispersion mode, 4) access to selected Zeeman-field modulation frequencies for optimizing sensitivity of ST-EPR measurements to changes in the rotational diffusion of a spin labeled protein and for analysis of data from multiple motional populations in ST-EPR experiments, and 5) support for maintenance and repair of the spectrometer and accessories. EPR spectroscopy is currently used by each of the applicants for studies which revolve around a central theme of structure-function relationships of proteins. This group has an established record of design and chemical synthesis of novel spin-label reagents for site-specific modification of proteins and for development of theoretical approaches for analysis of data obtained from well defined systems. The EPR work is complemented by a variety of other biophysical approaches including nuclear magnetic resonance (NMR), X-ray crystallography/molecular modeling, fluorescence spectroscopy, and molecular biology/protein engineering. The applicants have an established record of sharing instrumentation and of active collaborations in their research programs.