BIOPHYSICAL CHEMISTRY OF STEROID-PROTEIN INTERACTIONS

建议调查动态和静态过程在 类固醇和类固醇结合蛋白之间的相互作用。其目的是 为了更好地了解类固醇是如何结合的 它们的特殊性的基础。通过利用已知的光物理 使用稳定的非本征和本征发光探针的性质 状态和纳秒时间分辨荧光光谱，一种更 将开发关于类固醇结合部位和其 与蛋白质功能和结构的关系。这项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)，利用物种比较来帮助 阐明类固醇特异性的基础。远景目标是 了解类固醇如何与细胞内受体蛋白相互作用。这个 眼下的目标是了解血清蛋白质。将使用的方法 在这些研究中将有时间分辨和稳态的荧光和 类固醇结合试验。具体目标是： 1.马兜铃素与人结合的双态荧光衰减动力学 将对兔子的SBP进行研究，以提供有关该化学物质的信息 类固醇结合部位的环境及其与特异性结合部位的相互作用 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 将执行至SBP和CBG，以获取有关 两种蛋白质的流体力学形态和结构稳定性。 3.将研究类固醇荧光探针，可能具有 对SBP和CBG的结合亲和力，包括致癌芳香族化合物 据报道，水生动物具有温和的雌激素活性。 4.用稀土元素研究Ca~(++)与SBP的相互作用。 5.将研究蛋白质碳水化合物的作用，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.