These authors contributed equally to this work.
Methods Appl Fluoresc. 2016 Nov 22;4(4):042004. doi: 10.1088/2050-6120/4/4/042004.
Subdomains of macromolecules often undergo large orientation changes during their catalytic cycles that are essential for their activity. Tracking these rearrangements in real time opens a powerful window into the link between protein structure and functional output. Site-specific labeling of individual molecules with polarized optical probes and measurement of their spatial orientation can give insight into the crucial conformational changes, dynamics, and fluctuations of macromolecules. Here we describe the range of single molecule optical technologies that can extract orientation information from these probes, review the relevant types of probes and labeling techniques, and highlight the advantages and disadvantages of these technologies for addressing specific inquiries.
大分子的亚结构域在其催化循环中经常发生大的取向变化，这对它们的活性是必不可少的。实时跟踪这些重排为我们了解蛋白质结构与功能输出之间的联系提供了一个有力的窗口。用极化光学探针对单个分子进行特异性标记，并测量其空间取向，可以深入了解大分子的关键构象变化、动态和波动。在这里，我们描述了一系列可以从这些探针中提取取向信息的单分子光学技术，回顾了相关类型的探针和标记技术，并强调了这些技术在解决特定问题时的优缺点。
