Greenfeld Max, Herschlag Daniel
Department of Chemical Engineering, Stanford University, Stanford, California, USA.
Methods Enzymol. 2010;472:205-20. doi: 10.1016/S0076-6879(10)72009-7.
Tertiary contacts are critical to stabilizing the folded conformations of structured RNAs. In some cases, these contacts have been shown to interact with positive cooperativity. Measuring the energetic coupling of tertiary contact formation is among the most basic physical characterizations of a structured RNA. With proper experimental design, single-molecule fluorescence resonance energy transfer (smFRET) allows the rigorous determination of the energetic coupling. This chapter aims to provide a general experimental approach to measuring the energetic coupling of tertiary contacts, using smFRET.
三级相互作用对于稳定结构化RNA的折叠构象至关重要。在某些情况下，这些相互作用已被证明具有正协同性。测量三级相互作用形成的能量耦合是结构化RNA最基本的物理特性之一。通过适当的实验设计，单分子荧光共振能量转移（smFRET）可以精确测定能量耦合。本章旨在提供一种使用smFRET测量三级相互作用能量耦合的通用实验方法。
