Yang Byeongseon, Liu Zhaowei, Liu Haipei, Nash Michael A
Department of Chemistry, University of Basel, Basel, Switzerland.
Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland.
Front Mol Biosci. 2020 May 19;7:85. doi: 10.3389/fmolb.2020.00085. eCollection 2020.
Single-molecule force spectroscopy with the atomic force microscope provides molecular level insights into protein function, allowing researchers to reconstruct energy landscapes and understand functional mechanisms in biology. With steadily advancing methods, this technique has greatly accelerated our understanding of force transduction, mechanical deformation, and mechanostability within single- and multi-domain polyproteins, and receptor-ligand complexes. In this focused review, we summarize the state of the art in terms of methodology and highlight recent methodological improvements for AFM-SMFS experiments, including developments in surface chemistry, considerations for protein engineering, as well as theory and algorithms for data analysis. We hope that by condensing and disseminating these methods, they can assist the community in improving data yield, reliability, and throughput and thereby enhance the information that researchers can extract from such experiments. These leading edge methods for AFM-SMFS will serve as a groundwork for researchers cognizant of its current limitations who seek to improve the technique in the future for in-depth studies of molecular biomechanics.
利用原子力显微镜进行的单分子力谱技术能够在分子水平上深入了解蛋白质功能，使研究人员能够重建能量景观并理解生物学中的功能机制。随着方法的不断进步，该技术极大地加速了我们对单域和多域多聚蛋白以及受体-配体复合物内的力转导、机械变形和机械稳定性的理解。在这篇重点综述中，我们从方法学角度总结了当前的技术水平，并突出了原子力显微镜单分子力谱实验最近的方法改进，包括表面化学的进展、蛋白质工程的考量以及数据分析的理论和算法。我们希望通过浓缩和传播这些方法，能够帮助该领域提高数据产量、可靠性和通量，从而增强研究人员从此类实验中提取信息的能力。这些原子力显微镜单分子力谱的前沿方法将为那些认识到其当前局限性并寻求在未来改进该技术以深入研究分子生物力学的研究人员奠定基础。
