Zernike Institute for Advanced Materials, Centre for Synthetic Biology, University of Groningen, Nijenborgh 4, Groningen, The Netherlands.
Curr Opin Biotechnol. 2011 Feb;22(1):75-80. doi: 10.1016/j.copbio.2010.10.002. Epub 2010 Oct 29.
Single-molecule fluorescence techniques have emerged as powerful tools to study biological processes at the molecular level. This review describes the application of these methods to the characterization of the kinetics of interaction between biomolecules. A large number of single-molecule assays have been developed that visualize association and dissociation kinetics in vitro by fluorescently labeling binding partners and observing their interactions over time. Even though recent progress has been significant, there are certain limitations to this approach. To allow the observation of individual, fluorescently labeled molecules requires low, nanomolar concentrations. I will discuss how such concentration requirements in single-molecule experiments limit their applicability to investigate intermolecular interactions and how recent technical advances deal with this issue.
单分子荧光技术已成为研究分子水平生物过程的有力工具。本综述描述了这些方法在生物分子相互作用动力学特性分析中的应用。已经开发了大量的单分子检测方法，通过荧光标记结合配偶体并观察它们随时间的相互作用，在体外可视化结合和解离动力学。尽管最近取得了重大进展，但这种方法仍存在一定的局限性。为了能够观察到单个荧光标记的分子，需要使用低纳摩尔浓度的荧光标记物。我将讨论单分子实验中这种浓度要求如何限制其在研究分子间相互作用中的适用性，以及最近的技术进步如何解决这个问题。
