Department of Chemistry, University at Albany, State University of New York, 1400 Washington Ave, Albany, NY, 12222, USA.
Commun Biol. 2020 May 21;3(1):249. doi: 10.1038/s42003-020-0976-3.
Protein-protein interactions, PPIs, underlie most cellular processes, but many PPIs depend on a particular metabolic state that can only be observed in live, actively metabolizing cells. Real time in-cell NMR spectroscopy, RT-NMR, utilizes a bioreactor to maintain cells in an active metabolic state. Improvement in bioreactor technology maintains ATP levels at >95% for up to 24 hours, enabling protein overexpression and a previously undetected interaction between prokaryotic ubiquitin-like protein, Pup, and mycobacterial proteasomal ATPase, Mpa, to be detected. Singular value decomposition, SVD, of the NMR spectra collected over the course of Mpa overexpression easily identified the PPIs despite the large variation in background signals due to the highly active metabolome.
蛋白质-蛋白质相互作用（PPIs）是大多数细胞过程的基础，但许多 PPI 依赖于特定的代谢状态，而这种状态只能在活的、活跃代谢的细胞中观察到。实时细胞内 NMR 光谱学（RT-NMR）利用生物反应器使细胞保持活跃的代谢状态。生物反应器技术的改进使 ATP 水平保持在 >95%长达 24 小时，从而能够进行蛋白质过表达，并检测到先前未检测到的原核泛素样蛋白 Pup 和分枝杆菌蛋白酶体 ATP 酶 Mpa 之间的相互作用。尽管由于高度活跃的代谢组导致背景信号变化很大，但通过对 Mpa 过表达过程中收集的 NMR 光谱进行奇异值分解（SVD），很容易识别出 PPI。
