STOPPED-FLOW CD AND FLUORESCENCE SPECTROMETER

这项提案要求NCRR提供资金，用于购买应用光物理学pi*180 CDF光谱仪(APP)，以促进对生物大分子之间相互作用的动力学研究，以及对其构象稳定性的动力学和热力学研究。主要用户实验室的生化研究的一个主要目标是了解各种生化相互作用的分子本质，包括蛋白质-蛋白质相互作用、蛋白质-核酸相互作用以及蛋白质和核酸的折叠。其中许多研究包括对感兴趣的生物大分子中特定变化的详细结构-功能分析。这些相互作用在分子识别的基本生化过程中以及在不同的病理过程中都是重要的。所有这些现象都共享可检测到的光谱信号，这些信号要么对应于复杂的形成，要么对应于构象变化。这些过程通常发生在10毫秒之间的时间尺度上。还有一分钟叫L。这种时间制度需要一个特殊的样品处理装置，即停流装置，以产生必要的快速混合时间。APP光谱仪允许使用同一仪器同时监测圆二色谱、UV-VIS、荧光强度和荧光各向异性。这一关键特征允许直接比较光谱变化，便于将这些性质与特定的结构元素相关联。凯斯西部储备大学医学院致力于在分子生物学核心实验室的支持下提供必要的基础设施，并为仪器的长期维护做了准备。重要的是，CWRU园区目前没有提供与APP光谱仪相同功能的仪器。这对用户群体的研究计划构成了明显的限制。所要求的仪器将大大扩大主要用户资助的研究的范围，并将向大学社区提供访问ART动态测量网站的机会。

This proposal requests NCRR funds for the purchase of an Applied Photophysics pi* 180 CDF Spectrometer (APP) to facilitate the kinetic studies of the interactions between biological macromolecules, as well as kinetic and thermodynamic studies of their conformational stability. A major goal of biochemical research in the major users' laboratories is to understand the m9lecular nature of a variety of biochemical interactions including protein-protein interactions, protein-nucleic acid interactions, and the folding of both protein and nucleic acids. Many of these studies include detailed structure-function analyses of specific alterations in the biomacromolecules of interest. These interactions are important in both the fundamental biochemical processes of molecular recognition, as well as in different pathological processes. All of these phenomena share detectable spectroscopic signals that correspond either to complex formation, or to conformational change. These processes routinely occur on time scales of between 10 msec. and ca. l minute. This time regime requires a special sample handling unit, i.e. a stopped-flow, to produce the requisite rapid mixing times. The APP spectrometer permits circular dichroism, UV-Vis, fluorescence intensity and fluorescence anisotropy to be monitored simultaneously by the same instrument. This crucial feature permits the spectroscopic changes to be compared directly, facilitating the correlation of these properties to specific elements of structure. The Case Western Reserve University School of Medicine is committed to providing the necessary infrastructure under the auspices of the Molecular Biology Core Laboratory, and has made provisions for the long-term maintenance of instrument. Importantly, there are no instruments currently on the CWRU campus that provide the same capabilities as the APP spectrometer. This presents an obvious limitation to the research programs of the user group. The requested instrumentation will significantly enhance the scope of the major users' funded research and will provide access to site of the art kinetic measurements to the University community.