Advanced R&D Center, HORIBA, Ltd., 2 Miyanohigashi, Kisshoin, Minami-ku, Kyoto 601-8510, Japan.
Spectrochim Acta A Mol Biomol Spectrosc. 2017 Mar 15;175:145-154. doi: 10.1016/j.saa.2016.12.010. Epub 2016 Dec 15.
Investigation of the behaviour of proteins in crowded environments is crucial for understanding the role of proteins in biological environments. In this study, the behaviour of bovine serum albumin (BSA) in crowded (highly concentrated) environments was investigated using time-resolved fluorescence spectroscopy as a model system. By using energy transfer as a molecular ruler, the crowding effect was clearly observed in the time resolved spectra. In addition, by using both time resolved anisotropy measurement and Raman spectroscopy, more detail insights from conformational and dynamic points of view were described. Consequently, it was revealed that in the highly concentrated solution, most of the BSA molecules are in the fast-reversible oligomeric state and the association at the "hard" and "soft" interfaces between protein surfaces occurred in a highly crowded environment with the aid of a charge-charge and short-range attractive interface. From both the conformational and dynamic aspects, the detail spectroscopic understanding of the behaviour of BSA in the crowding environment was obtained.
研究蛋白质在拥挤环境中的行为对于理解蛋白质在生物环境中的作用至关重要。在本研究中，使用时间分辨荧光光谱法作为模型系统研究了牛血清白蛋白（BSA）在拥挤（高浓度）环境中的行为。通过将能量转移用作分子标尺，可以清楚地观察到时间分辨光谱中的拥挤效应。此外，通过使用时间分辨各向异性测量和拉曼光谱，从构象和动态角度描述了更详细的见解。因此，结果表明，在高浓度溶液中，大多数 BSA 分子处于快速可逆的寡聚状态，并且在电荷-电荷和短程吸引力界面的帮助下，在蛋白质表面之间的“硬”和“软”界面处发生缔合。从构象和动态两个方面，获得了 BSA 在拥挤环境中行为的详细光谱理解。
