Acquisition of an Argon Laser for Phase Fluorometry

获取用于相位荧光测定的氩激光器

• 批准号: 9120260
• 负责人: Steven Grossman
• 金额: $ 4.12万
• 依托单位: University of South Florida
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1993-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9120260&HistoricalAwards=false
• 关键词: Acquisition; Argon; Laser; Phase; Fluorometry

An argon laser will be used in fluorescence decay studies of creatine kinase, cascade polymers, nitrate reductase and an investigation of the static quenching limit. Intrinsic and extrinsic fluorescence lifetime and anisotropy investigations of creatine kinase folding and conformation are proposed. A comprehensive set of equilibrium and kinetic experiments, including quenching studies are proposed to describe intermediates in the folding pathway and the temporal relationship between folding and association of subunits. Experiments will assess the internal environment of cascade polymers in relation to synthetic micelles, using lipophilic probe lifetimes and anisotropies. Experiments are proposed to evaluate the solvent characteristics of the internal domain and the degree of packing using diphenylhexatriene, a well- characterized probe of micelle dynamics. A study is presented to characterize the relationship between catalytic properties and the conformations of the dimer, tetramer and isolated domains of nitrate reductase. Experiments are proposed to examine the assembly of the tetrameric state, using the intrinsic fluorescence and thiol-labelled protein as reporters of conformational changes in the presence of effectors of activity. Lifetime measurements will be used to obtain quenching constants for perylene and 9,10 dicyanoanthracence by carbon tetrachloride and used in calculation of the static quenching limit. The argon laser is a high intensity excitation source required for examining intrinsic fluorescence lifetimes and extrinsic fluorophore anisotropies. The multiple excitation wavelengths will be useful in global analysis of overdetermined data sets to characterize decay laws using discrete and distributed models.

氩激光器将用于荧光衰减研究 肌酸激酶，级联聚合物，硝酸还原酶和 静态猝灭极限的研究。 本征和非本征荧光寿命和各向异性 肌酸激酶折叠和构象的研究， 提出了 一套全面的平衡和动力学 实验，包括淬火研究提出了描述 折叠途径中的中间体和时间 折叠和亚基结合之间的关系。 实验将评估级联的内部环境 与合成胶束相关的聚合物，使用亲脂性 探针寿命和各向异性。 实验被提议为 评估内部域的溶剂特性， 使用二苯基己三烯的填充程度， 胶束动力学的表征探针。 一项研究提出了表征之间的关系 二聚体、四聚体的构象和催化性能 和硝酸还原酶的分离结构域。 实验 建议检查四聚体状态的组装，使用 本征荧光和巯基标记的蛋白质， 在效应物存在下的构象变化报告基因 的活动。 寿命测量将用于获得猝灭常数 用四氯化碳对苝和9，10-二氰基蒽进行了测定 并用于静态猝灭极限的计算。 氩激光器是所需的高强度激发源 用于检查固有荧光寿命和非固有荧光寿命， 荧光团各向异性。 多个激发波长 将有助于超定数据集的全局分析， 使用离散和分布式模型表征衰减规律。