STOPPED-FLOW CD AND FLUORESCENCE SPECTROMETER

停流 CD 和荧光光谱仪

• 批准号: 6062442
• 负责人: MICHAEL E. HARRIS
• 金额: $ 14.73万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6062442
• 关键词: biomedical equipment purchase; fluorescence spectrometry; stop flow technique

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.

本提案申请NCRR资金购买应用光物理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，以及其构象稳定性的动力学和热力学研究。在主要用户实验室中，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性质，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。许多这些研究包括对感兴趣的生物大分子的特定变化进行详细的结构-功能分析。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。所有这些现象都具有可检测的光谱信号，这些信号要么与复杂的形成相对应，要么与构象变化相对应。这些过程通常发生在10毫秒之间的时间尺度上。大约1分钟。这种时间制度需要一个特殊的样品处理单元，即停止流动，以产生所需的快速混合时间。APP光谱仪允许在同一台仪器上同时监测圆二色性、紫外可见性、荧光强度和荧光各向异性。这一关键特征使光谱变化可以直接进行比较，促进了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提供了规定。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。这对用户群的研究方案提出了明显的限制。所要求的仪器将大大扩大主要用户资助的研究范围，并将为大学社区提供艺术动力学测量的访问地点。