Müller Daniel J, Engel Andreas
Center of Biotechnology, University of Technology, Tatzberg 47-51, Dresden, Germany.
Nat Protoc. 2007;2(9):2191-7. doi: 10.1038/nprot.2007.309.
Membrane proteins comprise 30% of the proteome of higher organisms. They mediate energy conversion, signal transduction, solute transport and secretion. Their native environment is a bilayer in a physiological buffer solution, hence their structure and function are preferably assessed in this environment. The surface structure of single membrane proteins can be determined in buffer solutions by atomic force microscopy (AFM) at a lateral resolution of less than 1 nm and a vertical resolution of 0.1-0.2 nm. Moreover, single proteins can be directly addressed, stuck to the AFM stylus and subsequently unfolded, revealing the molecular interactions of the protein studied. The examples discussed here illustrate the power of AFM in the structural analysis of membrane proteins in a native environment.
膜蛋白占高等生物蛋白质组的30%。它们介导能量转换、信号转导、溶质转运和分泌。它们的天然环境是生理缓冲溶液中的双层膜,因此其结构和功能最好在这种环境中进行评估。单膜蛋白的表面结构可以在缓冲溶液中通过原子力显微镜(AFM)以小于1 nm的横向分辨率和0.1 - 0.2 nm的垂直分辨率来确定。此外,单个蛋白质可以直接被处理,附着在AFM探针上,随后展开,揭示所研究蛋白质的分子相互作用。这里讨论的例子说明了AFM在天然环境中膜蛋白结构分析中的作用。