PHOTOCHEMICAL PROBES OF PROTEIN FLEXIBILITY

蛋白质柔性的光化学探针

• 批准号: 3280592
• 负责人: ANGELO A LAMOLA
• 金额: $ 3.76万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1985-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3280592
• 关键词: apoenzymes; cis trans isomerization; myoglobin; nuclear magnetic resonance spectroscopy; photochemistry; stilbenes

The goal of the proposed research is to develop photochemical probes of globular protein structural fluctuation dynamics in the subnanosecond time regime. Specific aims include the delineation of the dynamics of the light-induced configurational (cis trans) isomerization of protein-bound stilbene and stilbene derivatives and of dipyrromethenone derivatives. Because rotation (leading to isomerization) in the excited singlet states of these compounds is fast (10-100 ps) and intersystem crossing is relatively slow, rotational dynamics can be related to fluorescence intensities and fluorescence lifetimes in simple ways. Rotational barriers provided by the protein environment can then be detected with facility. Effects of substituent size on isomerization dynamics, reflective of the "space" requirements for rotation, should yield additional information about the flexibility of the protein binding site. Effects of solvent viscosity on isomerization dynamics should relate to solvent protein interactions. Interpretation of dynamical data will require knowledge about the sites of binding of the probes to the protein and choice of proteins is critical. Proteins with "known" structures will be utilized. Apomyoglobin is planned for the first studies. A variety of approaches including high resolution nmr should reveal the binding sites. In more mature studies, derivatives of stilbene and dipyrromethenone that bind to active sites of enzymes (e.g., lysozyme) will be prepared to probe fast structural fluctuations in those sites and possible relations with enzymic function.

拟议的研究的目的是开发 球蛋白结构波动动力学 政权。 具体目的包括划定的动力学 光诱导的构型（顺式反式）异构化蛋白结合 Stilbene和Stilbene衍生物以及二吡俄替代衍生物。 因为旋转（导致异构化）在激发的单线状态下 这些化合物的快速（10-100 ps），并且系统交叉是 相对较慢，旋转动力学可能与荧光有关 强度和荧光寿命以简单的方式。 旋转障碍 然后可以用设施检测到蛋白质环境提供。 取代基尺寸对异构化动力学的影响，反映了 “空间”旋转要求应产生其他信息 关于蛋白质结合位点的灵活性。 溶剂的影响 异构化动力学上的粘度应与溶剂蛋白有关 互动。 动态数据的解释将需要知识 关于探针与蛋白质结合的位点和选择 蛋白质至关重要。 将使用具有“已知”结构的蛋白质。 计划进行初次研究。 多种方法 包括高分辨率NMR应揭示结合位点。 更多 成熟的研究，Stilbene和Dipyrromethenone的衍生物与 酶的活性位点（例如，溶菌酶）将准备快速探测 这些地点的结构波动以及可能与酶的关系 功能。