Thompson N L, Drake A W, Chen L, Vanden Broek W
Department of Chemistry, University of North Carolina, Chapel Hill 27599-3290, USA.
Photochem Photobiol. 1997 Jan;65(1):39-46. doi: 10.1111/j.1751-1097.1997.tb01875.x.
The equilibrium, kinetics, diffusion and self-association of proteins at membrane/solution interfaces may deviate substantially from these processes in bulk solution. A set of methods for examining these phenomena combines substrate-supported planar model membranes and the use of evanescent illumination with laser-based, quantitative fluorescence microscopy. Measurement of the steady-state, surface-associated fluorescence can be used to examine the thermodynamic properties of proteins at membranes. When combined with fluorescence photobleaching recovery, this technique provides information about membrane-binding kinetics; and when combined with fluorescence pattern photobleaching recovery, measurement of the translational diffusion coefficients of proteins weakly bound to membranes is possible. The use of polarized evanescent illumination can provide information about the orientation distributions of adsorbed fluorophores. Fluorescence correlation spectroscopy provides information about the self-association (e.g. dimerization) of membrane-associated proteins.
蛋白质在膜/溶液界面处的平衡、动力学、扩散和自缔合可能与本体溶液中的这些过程有很大偏差。一组用于研究这些现象的方法结合了底物支撑的平面模型膜以及利用倏逝光照明和基于激光的定量荧光显微镜。稳态表面相关荧光的测量可用于研究蛋白质在膜上的热力学性质。当与荧光光漂白恢复相结合时，该技术可提供有关膜结合动力学的信息；当与荧光图案光漂白恢复相结合时，则可以测量弱结合于膜的蛋白质的平移扩散系数。使用偏振倏逝光照明可提供有关吸附荧光团取向分布的信息。荧光相关光谱可提供有关膜相关蛋白质自缔合（例如二聚化）的信息。
