Acquisition of an Argon Laser for Phase Fluorometry

Ar激光将用于肌酸激酶、级联聚合物、硝酸还原酶的荧光衰减研究和静态猝灭极限的研究。对肌酸激酶折叠和构象的本征和外在荧光寿命以及各向异性进行了研究。提出了一套全面的平衡和动力学实验，包括猝灭研究，以描述折叠途径中的中间产物以及折叠和亚基结合之间的时间关系。实验将使用亲脂探针寿命和各向异性来评估与合成胶束相关的级联聚合物的内部环境。提出了使用二苯基己三烯的实验来评估内部区域的溶剂特性和堆积程度，二苯基己三烯是胶束动力学的良好表征的探针。研究了硝酸还原酶二聚体、四聚体和分离结构域的构象与催化性质的关系。实验被用来检测四聚体的组装，使用本征荧光和硫醇标记的蛋白质作为活性效应物存在时构象变化的报告。寿命测量将被用来获得茂和9，10-二氰基菲对四氯化碳的猝灭常数，并用于计算静态猝灭极限。Ar激光是一种高强度激发源，用于检测固有荧光寿命和外在荧光团各向异性。多个激发波长在超定数据集的全局分析中将是有用的，以使用离散和分布模型来表征衰减规律。

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.