Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zürich, Basel, Switzerland.
Wallenberg Centre for Molecular and Translational Medicine, Department of Chemistry and Molecular Biology, University of Gothenburg, Gothenburg, Sweden.
Methods Mol Biol. 2020;2127:359-372. doi: 10.1007/978-1-0716-0373-4_23.
Atomic force microscopy (AFM)-based single-molecule force spectroscopy allows direct physical manipulation of single membrane proteins under near-physiological conditions. It can be applied to study mechanical properties and molecular interactions as well as unfolding and folding pathways of membrane proteins. Here, we describe the basic procedure to study membrane proteins by single-molecule force spectroscopy and discuss general requirements of the experimental setup as well as common pitfalls typically encountered when working with membrane proteins in AFM.
原子力显微镜(AFM)基于单分子力谱技术可以在接近生理条件下直接对单个膜蛋白进行物理操作。它可以用于研究膜蛋白的机械性质和分子相互作用以及解折叠和折叠途径。在这里,我们描述了通过单分子力谱技术研究膜蛋白的基本步骤,并讨论了实验设置的一般要求以及在 AFM 中使用膜蛋白时通常遇到的常见问题。