School of Physics and Astronomy, University of Leeds, Leeds LS2 9JT, United Kingdom.
Chem Soc Rev. 2012 Jul 21;41(14):4781-96. doi: 10.1039/c2cs35033e. Epub 2012 May 30.
In recent years single molecule force spectroscopy has emerged as a powerful new tool to explore the mechanical stability and folding pathways of individual proteins. This technique is used to apply a stretching force between two points of a protein, unfolding the protein to an extended state. By measuring the unfolding and folding trajectories of individual proteins, insight can be gained into the physical mechanisms of protein folding. In this tutorial review we introduce the reader to single molecule force spectroscopy using the atomic force microscope (AFM), and explain the two main modes of operation of the AFM for force spectroscopy: force-extension and force-clamp. We introduce the approach of using polyproteins to obtain a clear mechanical fingerprint for monitoring the response of proteins to an applied mechanical force. In addition, we provide an informative and representative review of recent research on proteins using single molecule force spectroscopy. We focus on areas which have made a significant contribution to the single molecule protein folding field and highlight emerging areas of research which have wider implications for the general scientific community.
近年来，单分子力谱学已成为探索单个蛋白质机械稳定性和折叠途径的强大新工具。该技术用于在蛋白质的两点之间施加拉伸力，将蛋白质展开到伸展状态。通过测量单个蛋白质的展开和折叠轨迹，可以深入了解蛋白质折叠的物理机制。在本教程综述中，我们使用原子力显微镜 (AFM) 向读者介绍单分子力谱学，并解释 AFM 用于力谱学的两种主要操作模式：力-延伸和力-钳位。我们介绍了使用多蛋白的方法，以获得清晰的机械指纹，用于监测蛋白质对施加机械力的响应。此外，我们还对使用单分子力谱学研究蛋白质的最新研究进行了信息丰富且具有代表性的综述。我们重点介绍了对单分子蛋白质折叠领域做出重大贡献的领域，并强调了对更广泛科学界具有启示意义的新兴研究领域。
