Yamaoki Yudai, Nagata Takashi, Sakamoto Tomoki, Katahira Masato
Institute of Advanced Energy, Kyoto University, Uji, Kyoto 611-0011, Japan.
Graduate School of Energy Science, Kyoto University, Uji, Kyoto 611-0011, Japan.
Biophys Physicobiol. 2020 May 26;17:36-41. doi: 10.2142/biophysico.BSJ-2020006. eCollection 2020.
The intracellular environment is highly crowded with biomacromolecules such as proteins and nucleic acids. Under such conditions, the structural and biophysical features of nucleic acids have been thought to be different from those . To obtain high-resolution structural information on nucleic acids in living cells, the in-cell NMR method is a unique tool. Following the first in-cell NMR measurement of nucleic acids in 2009, several interesting insights were obtained using oocytes. However, the in-cell NMR spectrum of nucleic acids in living human cells was not reported until two years ago due to the technical challenges of delivering exogenous nucleic acids. We reported the first in-cell NMR spectra of nucleic acids in living human cells in 2018, where we applied a pore-forming toxic protein, streptolysin O. The in-cell NMR measurements demonstrated that the hairpin structures of nucleic acids can be detected in living human cells. In this review article, we summarize our recent work and discuss the future prospects of the in-cell NMR technique for nucleic acids.
细胞内环境中充斥着蛋白质和核酸等生物大分子。在这种情况下，核酸的结构和生物物理特性被认为与那些情况有所不同。为了获得活细胞中核酸的高分辨率结构信息，细胞内核磁共振方法是一种独特的工具。在2009年首次进行细胞内核酸核磁共振测量之后，利用卵母细胞获得了一些有趣的见解。然而，由于递送外源核酸的技术挑战，直到两年前才报道了活的人类细胞中核酸的细胞内核磁共振谱。我们在2018年报道了活的人类细胞中核酸的首个细胞内核磁共振谱，当时我们应用了一种成孔毒性蛋白——链球菌溶血素O。细胞内核磁共振测量表明，在活的人类细胞中可以检测到核酸的发夹结构。在这篇综述文章中，我们总结了我们最近的工作，并讨论了细胞内核酸核磁共振技术的未来前景。
