Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31 Biopolis way, Nanos, #03-01, Singapore 138669, Singapore.
Int J Mol Sci. 2019 Jan 2;20(1):139. doi: 10.3390/ijms20010139.
In-cell nuclear magnetic resonance (NMR) is a method to provide the structural information of a target at an atomic level under physiological conditions and a full view of the conformational changes of a protein caused by ligand binding, post-translational modifications or protein⁻protein interactions in living cells. Previous in-cell NMR studies have focused on proteins that were overexpressed in bacterial cells and isotopically labeled proteins injected into oocytes of or delivered into human cells. Applications of in-cell NMR in probing protein modifications, conformational changes and ligand bindings have been carried out in mammalian cells by monitoring isotopically labeled proteins overexpressed in living cells. The available protocols and successful examples encourage wide applications of this technique in different fields such as drug discovery. Despite the challenges in this method, progress has been made in recent years. In this review, applications of in-cell NMR are summarized. The successful applications of this method in mammalian and bacterial cells make it feasible to play important roles in drug discovery, especially in the step of target engagement.
细胞内核磁共振（NMR）是一种在生理条件下提供目标原子水平结构信息的方法，也是观察配体结合、翻译后修饰或蛋白质-蛋白质相互作用引起的蛋白质构象变化的全景的方法。以前的细胞内 NMR 研究主要集中在细菌细胞中过表达和同位素标记的蛋白质上，以及注入卵母细胞或递送到人类细胞中的蛋白质。通过监测活细胞中过表达的同位素标记蛋白质，在哺乳动物细胞中已经进行了细胞内 NMR 在探测蛋白质修饰、构象变化和配体结合中的应用。可用的方案和成功的例子鼓励该技术在不同领域（如药物发现）的广泛应用。尽管该方法存在挑战，但近年来已取得进展。在这篇综述中，总结了细胞内 NMR 的应用。该方法在哺乳动物和细菌细胞中的成功应用使得它在药物发现中特别是在靶标结合步骤中发挥重要作用成为可能。
