DNP-Supported Solid-State NMR Spectroscopy of Proteins Inside Mammalian Cells

被引：62
作者：
Narasimhan, Siddarth [1 ]
Scherpe, Stephan [2 ,3 ]
Paioni, Alessandra Lucini [1 ]
van der Zwan, Johan [1 ]
Folkers, Gert E. [1 ]
Ovaa, Huib [2 ,3 ]
Baldus, Marc [1 ]
机构：
[1] Univ Utrecht, NMR Spect Grp, Bijvoet Ctr Biomol Res, Padualaan 8, NL-3584 CH Utrecht, Netherlands
[2] LUMC, Oncode Inst, Einthovenweg 20, NL-2333 ZC Leiden, Netherlands
[3] LUMC, Dept Cell & Chem Biol, Einthovenweg 20, NL-2333 ZC Leiden, Netherlands
来源：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2019年 / 58卷 / 37期
基金：
欧盟地平线“2020”;
关键词：
dynamic nuclear polarization; in-cell NMR; protein-protein interactions; solid-state NMR; ubiquitination; DYNAMIC NUCLEAR-POLARIZATION; MEMBRANE-PROTEINS; UBIQUITIN; COMPLEX; TOOL;
D O I：
10.1002/anie.201903246
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
Elucidating at atomic level how proteins interact and are chemically modified in cells represents a leading frontier in structural biology. We have developed a tailored solid-state NMR spectroscopic approach that allows studying protein structure inside human cells at atomic level under high-sensitivity dynamic nuclear polarization (DNP) conditions. We demonstrate the method using ubiquitin (Ub), which is critically involved in cellular functioning. Our results pave the way for structural studies of larger proteins or protein complexes inside human cells, which have remained elusive to in-cell solution-state NMR spectroscopy due to molecular size limitations.
引用
收藏
页码：12969 / 12973
页数：5
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