Quantitative comparison of protein dynamics in live cells and in vitro by in-cell (19)F-NMR.
Quantitative comparison of protein dynamics in live cells and in vitro by in-cell (19)F-NMR.
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DOI：
10.1039/c3cc39205h
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发表时间：
2013-03
影响因子：
4.9
通讯作者：
Yousuke Takaoka;Y. Kioi;A. Morito;Junji Otani;K. Arita;E. Ashihara;M. Ariyoshi;H. Tochio;M. Shirakawa;I. Hamachi
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文献类型：
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作者：
Yousuke Takaoka;Y. Kioi;A. Morito;Junji Otani;K. Arita;E. Ashihara;M. Ariyoshi;H. Tochio;M. Shirakawa;I. Hamachi
在这里，我们描述了如何通过化学生物学方法将 (19)F 探针掺入内源蛋白质中，从而揭示蛋白质动力学。通过使用 X 射线晶体学明确测定配体结合和未结合的结构，定量比较活细胞和体外的蛋白质动力学。这些结果清楚地证明了细胞内蛋白质更大的构象波动，部分原因是大分子拥挤效应。
Here we describe how a (19)F-probe incorporated into an endogenous protein by a chemical biology method revealed protein dynamics. By explicit determination of ligand-bound and unbound structures with X-ray crystallography, the quantitative comparison of the protein's dynamics in live cells and in vitro is presented. These results clearly demonstrated the greater conformational fluctuations of the intracellular protein, partially due to macromolecular crowding effects.