Institute for Physical and Theoretical Chemistry, and Braunschweig Institute for Integrated Systems Biology (BRICS), and Laboratory for Emerging Nanometrology (LENA), TU Braunschweig, Rebenring 56, 38106 Braunschweig, Germany.
Nanoscale. 2018 Sep 13;10(35):16416-16424. doi: 10.1039/c8nr03361g.
Fluorescence resonance energy transfer (FRET) has been instrumental in determining the structure and dynamics of biomolecules but distances above 8 nanometers are not accessible. However, with the advent and rapid development of super-resolution (SR) microscopy, distances between two fluorescent dyes below 20 nanometers can be resolved, which hitherto has been inaccessible for fluorescence microscopy approaches due to the limited resolving power of an optical imaging system that is determined by the fundamental laws of light diffraction (referred to as the diffraction limit). Therefore, the question arises whether SR microscopy can ultimately close the resolution gap between FRET and the diffraction limit and whether SR microscopy can be employed for the structural interrogation of proteins in the sub-20 nm range? Here, we show that the combination of DNA nanotechnology and single-molecule biochemistry allows the first step towards the investigation of the structural organization of a protein via SR microscopy. Limiting factors and possible future directions for the full implementation of SR microscopy as a structural tool are discussed.
荧光共振能量转移（FRET）在确定生物分子的结构和动态方面发挥了重要作用，但距离大于 8 纳米的范围无法探测。然而，随着超分辨率（SR）显微镜的出现和快速发展，两个荧光染料之间的距离小于 20 纳米的范围可以被解析，这在以前由于光学成像系统的分辨率有限，而无法通过荧光显微镜方法实现，因为光学成像系统的分辨率是由光衍射的基本定律决定的（称为衍射极限）。因此，出现了一个问题，即 SR 显微镜是否最终能够缩小 FRET 和衍射极限之间的分辨率差距，以及 SR 显微镜是否可以用于在亚 20 纳米范围内对蛋白质进行结构检测？在这里，我们展示了 DNA 纳米技术和单分子生物化学的结合，为通过 SR 显微镜研究蛋白质的结构组织迈出了第一步。讨论了作为结构工具全面实施 SR 显微镜的限制因素和可能的未来方向。
