Müller Daniel J, Sapra K Tanuj, Scheuring Simon, Kedrov Alexej, Frederix Patrick L, Fotiadis Dimitrios, Engel Andreas
Center for Biotechnology, University of Technology, 01307 Dresden, Germany.
Curr Opin Struct Biol. 2006 Aug;16(4):489-95. doi: 10.1016/j.sbi.2006.06.001. Epub 2006 Jun 23.
Characterizing membrane proteins with single-molecule techniques provides structural and functional insights that cannot be obtained with conventional approaches. Recent studies show that atomic force microscopy (AFM) in the context of a 'lab on a tip' enables the measurement of multiple parameters of membrane proteins. This multifunctional tool can be applied to probe the oligomeric states and conformational changes of membrane protein assemblies in their native environment. The ability to determine diverse properties at high spatial resolution facilitates the mapping of structural flexibilities, electrostatic potentials and electric currents. By using the AFM tip as tweezer, it is possible to characterize unfolding and refolding pathways of single proteins and the location of their molecular interactions. These interactions dictate the stability of the protein and might be modulated by ligands that alter the protein's functional state.
用单分子技术表征膜蛋白可提供传统方法无法获得的结构和功能见解。最近的研究表明，“芯片实验室”背景下的原子力显微镜（AFM）能够测量膜蛋白的多个参数。这种多功能工具可用于探测天然环境中膜蛋白组装体的寡聚状态和构象变化。在高空间分辨率下确定多种特性的能力有助于绘制结构柔韧性、静电势和电流图。通过将AFM探针用作镊子，可以表征单个蛋白质的解折叠和重折叠途径及其分子相互作用的位置。这些相互作用决定了蛋白质的稳定性，并且可能受到改变蛋白质功能状态的配体的调节。
