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Laboratory diffracted x-ray blinking to monitor picometer motions of protein molecules and application to crystalline materials
Structural Dynamics ( IF 3.670 ) Pub Date : 2021-07-08 , DOI: 10.1063/4.0000112
Tatsuya Arai 1, 2 , Rena Inamasu 3 , Hiroki Yamaguchi 3 , Daisuke Sasaki 1 , Ayana Sato-Tomita 4 , Hiroshi Sekiguchi 5 , Kazuhiro Mio 2 , Sakae Tsuda 2, 6, 7 , Masahiro Kuramochi 1, 2 , Yuji C. Sasaki 1, 2, 5
Affiliation  

In recent years, real-time observations of molecules have been required to understand their behavior and function. To date, we have reported two different time-resolved observation methods: diffracted x-ray tracking and diffracted x-ray blinking (DXB). The former monitors the motion of diffracted spots derived from nanocrystals labeled onto target molecules, and the latter measures the fluctuation of the diffraction intensity that is highly correlated with the target molecular motion. However, these reports use a synchrotron x-ray source because of its high average flux, resulting in a high time resolution. Here, we used a laboratory x-ray source and DXB to measure the internal molecular dynamics of three different systems. The samples studied were bovine serum albumin (BSA) pinned onto a substrate, antifreeze protein (AFP) crystallized as a single crystal, and poly{2-(perfluorooctyl)ethyl acrylate} (PC8FA) polymer between polyimide sheets. It was found that not only BSA but also AFP and PC8FA molecules move in the systems. In addition, the molecular motion of AFP molecules was observed to increase with decreasing temperature. The rotational diffusion coefficients (DR) of BSA, AFP, and PC8FA were estimated to be 0.73 pm2/s, 0.65 pm2/s, and 3.29 pm2/s, respectively. Surprisingly, the DR of the PC8FA polymer was found to be the highest among the three samples. This is the first report that measures the molecular motion of a single protein crystal and polymer by using DXB with a laboratory x-ray source. This technique can be applied to any kind of crystal and crystalline polymer and provides atomic-order molecular information.

中文翻译:

实验室衍射 X 射线闪烁监测蛋白质分子的皮米运动以及在晶体材料中的应用

近年来,需要实时观察分子以了解其行为和功能。迄今为止,我们已经报道了两种不同的时间分辨观察方法:衍射 X 射线跟踪和衍射 X 射线闪烁 (DXB)。前者监测来自标记到目标分子上的纳米晶体的衍射点的运动,后者测量与目标分子运动高度相关的衍射强度的波动。然而,这些报告使用同步加速器 X 射线源,因为它的平均通量高,因此时间分辨率高。在这里,我们使用实验室 X 射线源和 DXB 来测量三个不同系统的内部分子动力学。研究的样品是固定在基质上的牛血清白蛋白 (BSA),8 FA) 聚酰亚胺片之间的聚合物。发现不仅BSA而且AFP和PC 8 FA分子在系统中移动。此外,观察到 AFP 分子的分子运动随着温度的降低而增加。BSA、AFP 和 PC 8 FA的旋转扩散系数 (D R )估计分别为 0.73 pm 2 /s、0.65 pm 2 /s 和 3.29 pm 2 /s。令人惊讶的是,PC 8的 D R发现 FA 聚合物在三个样品中最高。这是第一份通过实验室 X 射线源使用 DXB 测量单个蛋白质晶体和聚合物分子运动的报告。该技术可应用于任何类型的晶体和结晶聚合物,并提供原子级分子信息。
更新日期:2021-08-31
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