细胞内核磁共振谱的灵敏度和分辨率得到提高。
Improved sensitivity and resolution of in-cell NMR spectra.
作者信息
Burz David S, Breindel Leonard, Shekhtman Alexander
机构信息
Department of Chemistry, University at Albany, State University of New York, Albany, NY, United States.
Department of Chemistry, University at Albany, State University of New York, Albany, NY, United States.
出版信息
Methods Enzymol. 2019;621:305-328. doi: 10.1016/bs.mie.2019.02.029. Epub 2019 Mar 13.
In-cell NMR spectroscopy is a powerful tool to study protein structures and interactions under near physiological conditions in both prokaryotic and eukaryotic living cells. The low sensitivity and resolution of in-cell NMR spectra and limited lifetime of cells over the course of an in-cell experiment have presented major hurdles to wide acceptance of the technique, limiting it to a few select systems. These issues are addressed by introducing the use of the CRINEPT pulse sequence to increase the sensitivity and resolution of in-cell NMR spectra and the use of a bioreactor to maintain cell viability for up to 24h. Application of advanced pulse sequences and bioreactor during in-cell NMR experiments will facilitate the exploration of a wide range of biological processes.
细胞内核磁共振光谱是一种强大的工具，可用于研究原核生物和真核生物活细胞中接近生理条件下的蛋白质结构和相互作用。细胞内核磁共振光谱的低灵敏度和分辨率以及在细胞内实验过程中细胞的有限寿命，是该技术广泛应用面临的主要障碍，使其仅限于少数特定系统。通过引入CRINEPT脉冲序列来提高细胞内核磁共振光谱的灵敏度和分辨率，以及使用生物反应器将细胞活力维持长达24小时，这些问题得到了解决。在细胞内核磁共振实验中应用先进的脉冲序列和生物反应器将有助于探索广泛的生物过程。
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