Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, 119071, Russia.
Biochemistry (Mosc). 2012 Dec;77(13):1536-52. doi: 10.1134/S000629791213010X.
Atomic force microscopy (AFM) can be used to make measurements in vacuum, air, and water. The method is able to gather information about intermolecular interaction forces at the level of single molecules. This review encompasses experimental and theoretical data on the characterization of ligand-receptor interactions by AFM. The advantage of AFM in comparison with other methods developed for the characterization of single molecular interactions is its ability to estimate not only rupture forces, but also thermodynamic and kinetic parameters of the rupture of a complex. The specific features of force spectroscopy applied to ligand-receptor interactions are examined in this review from the stage of the modification of the substrate and the cantilever up to the processing and interpretation of the data. We show the specificities of the statistical analysis of the array of data based on the results of AFM measurements, and we discuss transformation of data into thermodynamic and kinetic parameters (kinetic dissociation constant, Gibbs free energy, enthalpy, and entropy). Particular attention is paid to the study of polyvalent interactions, where the definition of the constants is hampered due to the complex stoichiometry of the reactions.
原子力显微镜(AFM)可用于在真空、空气和水中进行测量。该方法能够在单分子水平上收集关于分子间相互作用力的信息。本综述涵盖了通过 AFM 表征配体-受体相互作用的实验和理论数据。与为表征单分子相互作用而开发的其他方法相比,AFM 的优势在于它不仅能够估计断裂力,还能够估计复合物断裂的热力学和动力学参数。本综述从基底和悬臂的修饰阶段一直到数据的处理和解释,检查了力谱学在配体-受体相互作用中的应用的具体特点。我们展示了基于 AFM 测量结果的基于数据的统计分析的特异性,并讨论了将数据转换为热力学和动力学参数(动力学解离常数、吉布斯自由能、焓和熵)。特别关注多价相互作用的研究,由于反应的复杂化学计量,常数的定义受到阻碍。