Institute for Molecules and Materials, Radboud University Nijmegen, The Netherlands.
Anal Biochem. 2011 Jul 1;414(1):1-6. doi: 10.1016/j.ab.2011.03.012. Epub 2011 Mar 12.
Single-molecule force spectroscopy (SMFS) using the atomic force microscope (AFM) has emerged as an important tool for probing biomolecular interaction and exploring the forces, dynamics, and energy landscapes that underlie function and specificity of molecular interaction. These studies require attaching biomolecules on solid supports and AFM tips to measure unbinding forces between individual binding partners. Herein we describe efficient and robust protocols for probing RNA interaction by AFM and show their value on two well-known RNA regulators, the Rev-responsive element (RRE) from the HIV-1 genome and an adenine-sensing riboswitch. The results show the great potential of AFM-SMFS in the investigation of RNA molecular interactions, which will contribute to the development of bionanodevices sensing single RNA molecules.
使用原子力显微镜（AFM）的单分子力谱学（SMFS）已成为探测生物分子相互作用以及探索分子相互作用的功能和特异性的基础的力、动力学和能量景观的重要工具。这些研究需要将生物分子附着在固体载体和 AFM 尖端上，以测量单个结合伴侣之间的解结合力。本文描述了通过 AFM 探测 RNA 相互作用的高效且稳健的方案，并展示了它们在两个著名的 RNA 调节剂上的价值，即 HIV-1 基因组中的 Rev 反应元件（RRE）和腺嘌呤感应核酶。结果表明，AFM-SMFS 在 RNA 分子相互作用研究方面具有巨大的潜力，这将有助于开发用于感应单个 RNA 分子的生物纳米器件。
