Evans E
Physics and Pathology, University of British Columbia, Vancouver, V6T 2A6, Canada.
Annu Rev Biophys Biomol Struct. 2001;30:105-28. doi: 10.1146/annurev.biophys.30.1.105.
On laboratory time scales, the energy landscape of a weak bond along a dissociation pathway is fully explored through Brownian-thermal excitations, and energy barriers become encoded in a dissociation time that varies with applied force. Probed with ramps of force over an enormous range of rates (force/time), this kinetic profile is transformed into a dynamic spectrum of bond rupture force as a function of loading rate. On a logarithmic scale in loading rate, the force spectrum provides an easy-to-read map of the prominent energy barriers traversed along the force-driven pathway and exposes the differences in energy between barriers. In this way, the method of dynamic force spectroscopy (DFS) is being used to probe the complex relation between force-lifetime-and chemistry in single molecular bonds. Most important, DFS probes the inner world of molecular interactions to reveal barriers that are difficult or impossible to detect in assays of near equilibrium dissociation but that determine bond lifetime and strength under rapid detachment. To use an ultrasensitive force probe as a spectroscopic tool, we need to understand the physics of bond dissociation under force, the impact of experimental technique on the measurement of detachment force (bond strength), the consequences of complex interactions in macromolecular bonds, and effects of multiply-bonded attachments.
在实验室时间尺度上,沿着解离路径的弱键能量景观通过布朗热激发得到充分探索,并且能垒被编码在随施加力而变化的解离时间中。在极宽的速率范围(力/时间)内用斜坡力进行探测,这种动力学轮廓会转变为作为加载速率函数的键断裂力动态谱。在加载速率的对数尺度上,力谱提供了一张易于读取的地图,展示了沿着力驱动路径所跨越的显著能垒,并揭示了能垒之间的能量差异。通过这种方式,动态力谱学(DFS)方法被用于探究单分子键中力 - 寿命 - 化学之间的复杂关系。最重要的是,DFS探测分子相互作用的内部世界,以揭示在近平衡解离测定中难以或无法检测到但在快速分离下决定键寿命和强度的能垒。要将超灵敏力探针用作光谱工具,我们需要了解力作用下键解离的物理过程、实验技术对分离力(键强度)测量的影响、大分子键中复杂相互作用的后果以及多键连接的影响。