BIOPHYSICAL CHEMISTRY OF STEROID-PROTEIN INTERACTIONS

该建议是调查动态和静态过程中， 类固醇和类固醇结合蛋白之间的相互作用。 目的是 为了更好地了解类固醇是如何结合的， 其特殊性的基础。 通过利用已知的非物理 非本征和本征发光探针的性质，使用稳态 状态和纳秒时间分辨荧光光谱，一个更 详细的图片将制定有关类固醇结合位点及其 与蛋白质功能和结构的关系。 工作将集中在两个方面 血清蛋白、性类固醇结合蛋白（SBP）和 皮质类固醇结合球蛋白（CBG），使用种属比较，以帮助 阐明类固醇特异性的基础。 长期目标是 了解类固醇如何与细胞内受体蛋白相互作用。 的 当前的目标是了解血清蛋白。 应采用的方法 在这些研究中，将是时间分辨和稳态荧光， 类固醇结合测定。 具体目标是： 1.马钱素与人结合的双态荧光衰减动力学 和兔SBP将进行研究，以提供有关化学品的信息 类固醇结合位点的环境和与特异性 个氨基酸残基 2.结合探针的时间分辨荧光各向异性衰减测量 将进行SBP和CBG，以获得有关 两种蛋白质的流体动力学形状和结构稳定性。 3.将研究类固醇样荧光探针，其可能具有 SBP和CBG的结合亲和力，包括致癌芳香族化合物 据报道，氢化物具有轻度雌激素活性。 4.将使用镧系元素研究Ca++与SBP的相互作用。 5.蛋白质碳水化合物的作用将被研究，以了解如何 它影响SBP和CBG的结构完整性和类固醇结合。

The proposal is to investigate dynamic and static processes in the interaction between steroids and steroid-binding proteins. The purpose is to develop a better understanding of how the steroids are bound and the basis for their specificity. By taking advantage of known photophysical properties of extrinsic and intrinsic luminescent probes, using steady state and nanosecond time-resolved fluorescence spectroscopy, a more detailed picture will be developed about the steroid binding site and its relationship to protein function and structure. The work will focus on two serum proteins, sex steroid-binding protein (SBP) and corticosteroid-binding globulin (CBG), using species comparison to help elucidate the basis for steroid specificity. The long range goal is to find out how steroids interact with intra-cellular receptor proteins. The immediate goal is to understand the serum proteins. The methods to be used in these studies will be time-resolved and steady state fluorescence and steroid binding assays. The specific aims are: 1. The two-state fluorescence decay kinetics of equilenin bound to human and rabbit SBP will be studied to provide information about the chemical environment of the steroid binding site and interactions with specific amino acid residues. 2. Time-resolved fluorescence anisotropy decay measurements of probes bound to SBP and CBG will be carried out to obtain information about the hydrodynamic shape and structural stability of the two proteins. 3. Steroid-like fluorescent probes will be studied which potentially have binding affinity for SBP and CBG, including carcinogenic aromatic hydrocarons reported to have mild estrogenic activity. 4. The interaction of Ca++ with SBP will be studied using lanthanides. 5. The role of the protein carbohydrate will be investigated to learn how it affects the structural integrity and steroid binding of SBP and CBG.