In-cell NMR: a topical review.
In-cell NMR: a topical review.
复制标题
DOI：
10.1107/s2052252516020625
复制
发表时间：
2017-03-01
期刊：
影响因子：
3.9
通讯作者：
Banci L
中科院分区：
文献类型：
--
作者：
Luchinat E;Banci L
细胞内核磁共振技术是近十年来发展起来的一种以原子级分辨率获取生物大分子结构和功能信息的新技术。本文综述了细胞内核磁共振技术的主要进展，重点介绍了细胞内核磁共振技术在真核生物和哺乳动物细胞中的应用。经典的结构生物学方法允许在体外对生物大分子进行结构表征，远离其生理背景。如今，由于丰富的结构数据以及技术和方法的进步，研究界的兴趣正逐渐从纯粹的结构测定转向生物分子功能方面的研究。因此，理想情况下需要细胞结构方法来表征生物分子，例如蛋白质，在其天然细胞环境中及其参与的功能过程。细胞内核磁共振是高分辨率核磁共振光谱的一种新应用，它可以直接在活细胞中分析蛋白质和其他大分子的结构和动力学特征。由于其挑战性，这一方法在过去15年中缓慢但稳步发展。迄今为止，几种细胞内NMR方法已成功应用于细菌和真核细胞，包括几种人类细胞系，并阐明了重要的结构和功能方面。本文综述了细胞内核磁共振技术的主要进展，重点介绍了细胞内核磁共振技术在真核生物和哺乳动物细胞中的应用。
In-cell NMR has emerged in the past decade as a unique approach to obtain structural and functional information on biological macromolecules within living cells at atomic resolution. In this review, the major advances of in-cell NMR are discussed, with a special focus on recent developments and applications in eukaryotic and mammalian cells. Classical structural biology approaches allow structural characterization of biological macromolecules in vitro, far from their physiological context. Nowadays, thanks to the wealth of structural data available and to technological and methodological advances, the interest of the research community is gradually shifting from pure structural determination towards the study of functional aspects of biomolecules. Therefore, a cellular structural approach is ideally needed to characterize biological molecules, such as proteins, in their native cellular environment and the functional processes that they are involved in. In-cell NMR is a new application of high-resolution nuclear magnetic resonance spectroscopy that allows structural and dynamical features of proteins and other macromolecules to be analyzed directly in living cells. Owing to its challenging nature, this methodology has shown slow, but steady, development over the past 15 years. To date, several in-cell NMR approaches have been successfully applied to both bacterial and eukaryotic cells, including several human cell lines, and important structural and functional aspects have been elucidated. In this topical review, the major advances of in-cell NMR are summarized, with a special focus on recent developments in eukaryotic and mammalian cells.