STOPPED-FLOW CD AND FLUORESCENCE SPECTROMETER

该提案要求NCRR提供资金，用于购买应用光物理学pi* 180 CDF光谱仪（APP），以促进生物大分子之间相互作用的动力学研究，以及构象稳定性的动力学和热力学研究。主要用户实验室中的生物化学研究的主要目标是了解各种生物化学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。许多这些研究包括详细的结构-功能分析的特定改变的生物大分子的利益。这些相互作用在分子识别的基本生化过程以及不同的病理过程中都很重要。所有这些现象共享可检测的光谱信号，其对应于复合物形成或构象变化。这些过程通常发生在10毫秒之间的时间尺度上。和约我一分钟。该时间方案需要特殊的样品处理单元，即停流，以产生所需的快速混合时间。APP光谱仪允许圆二色性、紫外-可见光、荧光强度和荧光各向异性由同一仪器同时监测。这一关键特征允许直接比较光谱变化，促进这些性质与特定结构元素的相关性。凯斯西储大学医学院致力于在分子生物学核心实验室的主持下提供必要的基础设施，并为仪器的长期维护做出了规定。重要的是，目前CWRU校园内没有仪器提供与APP光谱仪相同的功能。这对用户组的研究计划提出了明显的限制。所要求的仪器将大大提高主要用户资助的研究范围，并将提供访问网站的艺术动力学测量大学社区。

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.