Magnetic Resonance Center, CERM, University of Florence, 50019, Sesto Fiorentino, Italy.
Aix Marseille Univ, CNRS, ICR, Institut de Chimie Radicalaire, 13013, Marseille, France.
Chembiochem. 2020 Feb 17;21(4):451-460. doi: 10.1002/cbic.201900291. Epub 2019 Sep 26.
Exploring the structure and dynamics of biomolecules in the context of their intracellular environment has become the ultimate challenge for structural biology. As the cellular environment is barely reproducible in vitro, investigation of biomolecules directly inside cells has attracted a growing interest. Among magnetic resonance approaches, site-directed spin labeling (SDSL) coupled to electron paramagnetic resonance (EPR) spectroscopy provides competitive and advantageous features to capture protein structure and dynamics inside cells. To date, several in-cell EPR approaches have been successfully applied to both bacterial and eukaryotic cells. In this review, the major advances of in-cell EPR spectroscopy are summarized, as well as the challenges this approach still poses.
探索生物分子在细胞内环境中的结构和动态已成为结构生物学的终极挑战。由于细胞环境在体外几乎无法重现，因此直接在细胞内研究生物分子引起了越来越多的关注。在磁共振方法中，与电子顺磁共振（EPR）光谱学相结合的定点自旋标记（SDSL）提供了捕获细胞内蛋白质结构和动态的竞争优势。迄今为止，几种细胞内 EPR 方法已成功应用于细菌和真核细胞。本文总结了细胞内 EPR 光谱学的主要进展，以及该方法仍然面临的挑战。
