Diffracted X-ray Blinking Tracks Single Protein Motions.
Diffracted X-ray Blinking Tracks Single Protein Motions.
复制标题
衍射的X射线闪烁轨迹轨迹单蛋白运动。
DOI：
10.1038/s41598-018-35468-3
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发表时间：
2018-11-30
影响因子：
4.6
通讯作者：
Sasaki YC
中科院分区：
文献类型：
--
作者：
Sekiguchi H;Kuramochi M;Ikezaki K;Okamura Y;Yoshimura K;Matsubara K;Chang JW;Ohta N;Kubo T;Mio K;Suzuki Y;Chavas LMG;Sasaki YC
单分子动力学研究已经开始使用量子探针。使用低温透射电子显微镜的单粒子分析在研究蛋白质结构时显著提高了分辨率，并正在转向分子运动观察。X射线自由电子激光器也被探索作为确定生物实体的单分子结构的途径。在这里，我们提出了一种新的X射线单分子技术，可以在很长的时间尺度上观察分子的内部运动，从毫秒到103秒。我们的方法使用低剂量单色X射线和荧光标记技术。在单色X射线衍射实验中，由于水溶液中的布朗运动，移动单个纳米晶体的X射线衍射强度似乎会闪烁。观察到来自移动纳米晶体的X射线衍射斑点在布拉格条件下循环进出。因此，用纳米晶体标记的蛋白质分子的内部运动可以使用这种衍射X射线闪烁（DXB）方法从时间轨迹中提取。最后，我们成功地区分的程度的波动运动的一个单独的乙酰胆碱结合蛋白（AChBP）与乙酰胆碱（ACh）相互作用，使用实验室X射线源。
Single molecule dynamics studies have begun to use quantum probes. Single particle analysis using cryo-transmission electron microscopy has dramatically improved the resolution when studying protein structures and is shifting towards molecular motion observations. X-ray free-electron lasers are also being explored as routes for determining single molecule structures of biological entities. Here, we propose a new X-ray single molecule technology that allows observation of molecular internal motion over long time scales, ranging from milliseconds up to 103 seconds. Our method uses both low-dose monochromatic X-rays and nanocrystal labelling technology. During monochromatic X-ray diffraction experiments, the intensity of X-ray diffraction from moving single nanocrystals appears to blink because of Brownian motion in aqueous solutions. X-ray diffraction spots from moving nanocrystals were observed to cycle in and out of the Bragg condition. Consequently, the internal motions of a protein molecule labelled with nanocrystals could be extracted from the time trajectory using this diffracted X-ray blinking (DXB) approach. Finally, we succeeded in distinguishing the degree of fluctuation motions of an individual acetylcholine-binding protein (AChBP) interacting with acetylcholine (ACh) using a laboratory X-ray source.
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影响因子：
4.6
作者：
Sekiguchi H;Suzuki Y;Nishino Y;Kobayashi S;Shimoyama Y;Cai W;Nagata K;Okada M;Ichiyanagi K;Ohta N;Yagi N;Miyazawa A;Kubo T;Sasaki YC
通讯作者：
Sasaki YC
影响因子：
64.8
作者：
Sterling, Alphonse C.;Moore, Ronald L.;Adams, Mitzi
通讯作者：
Adams, Mitzi
影响因子：
6.8
作者：
Klepeis, John L.;Lindorff-Larsen, Kresten;Shaw, David E.
通讯作者：
Shaw, David E.
影响因子：
2.4
作者：
Okumura, Y;Oka, T;Sasaki, YC
通讯作者：
Sasaki, YC
影响因子：
2.5
作者：
Shinohara, Yuya;Watanabe, Akira;Amemiya, Yoshiyuki
通讯作者：
Amemiya, Yoshiyuki