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首页> 外文会议>Ultrasensitive and Single-Molecule Detection Technologies II; Progress in Biomedical Optics and Imaging; vol.8 no.21; Proceedings of SPIE-The International Society for Optical Engineering; vol.6444 >Monitoring of small conformational changes by high-precision measurements of hydrodynamic radius with 2-focus fluorescence correlation spectroscopy (2fFCS)
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Monitoring of small conformational changes by high-precision measurements of hydrodynamic radius with 2-focus fluorescence correlation spectroscopy (2fFCS)

机译：通过2焦点荧光相关光谱法（2fFCS）高精度测量流体动力学半径来监测小的构象变化

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摘要

We report on our application of a new fluorescence-correlation spectroscopy technique, 2-focus FCS, for measuring the hydrodynamic radius of molecules with sub- Angstrom precision. The method is applied of monitoring conformational changes of proteins upon ion binding. In particular, we present measurements on Ca~(2+)-binding of recoverin. Recoverin belongs to the superfamily of EF-hand Ca~(2+)-binding proteins and operates as a Ca~(2+)-sensor in vertebrate photoreceptor cells, where it regulates the activity of rhodopsin kinase GRK1 in a Ca~(2+)-dependent manner. The protein undergoes conformational changes upon Ca~(2+)-changes that are reflected as changes in their hydrodynamic radius. By using 2fFCS we were able to resolve hydrodynamic radius changes of ca. one Angstrom and used the Ca~(2+) dependence of this radius for recording binding curves in solution. We compare our results with those obtained by other techniques.
机译：我们报告了我们的新型荧光相关光谱技术，即2-焦点FCS，用于以亚埃精度测量分子的流体动力学半径。该方法用于监测离子结合后蛋白质的构象变化。特别地，我们提出了关于recoverin的Ca〜（2+）结合的测量。 Recoverin属于EF手Ca〜（2+）结合蛋白的超家族，在脊椎动物感光细胞中起Ca〜（2+）传感器的作用，调节着Ca〜（2）中视紫红质激酶GRK1的活性。 +）依赖方式。蛋白质在Ca〜（2 +）-变化时经历构象变化，反映为它们的流体动力学半径的变化。通过使用2fFCS，我们能够解析出ca的水动力半径变化。 1埃，并使用该半径的Ca〜（2+）依赖性记录溶液中的结合曲线。我们将我们的结果与通过其他技术获得的结果进行比较。

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