The room temperature crystal structure of a bacterial phytochrome determined by serial femtosecond crystallography.

The room temperature crystal structure of a bacterial phytochrome determined by serial femtosecond crystallography.
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DOI:
10.1038/srep35279
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发表时间:
2016-10-19
期刊:
影响因子:
4.6
通讯作者:
Westenhoff S
Westenhoff S
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Edlund P;Takala H;Claesson E;Henry L;Dods R;Lehtivuori H;Panman M;Pande K;White T;Nakane T;Berntsson O;Gustavsson E;Båth P;Modi V;Roy-Chowdhury S;Zook J;Berntsen P;Pandey S;Poudyal I;Tenboer J;Kupitz C;Barty A;Fromme P;Koralek JD;Tanaka T;Spence J;Liang M;Hunter MS;Boutet S;Nango E;Moffat K;Groenhof G;Ihalainen J;Stojković EA;Schmidt M;Westenhoff S

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光敏色素是一类控制植物、真菌和细菌对光反应的光感受器。一系列尚未完全理解的结构变化导致输出结构域的激活。时间分辨连续飞秒晶体学(SFX)可以潜在地照亮这些构象变化。在这里,我们报告的发色团结合结构域的耐辐射异常球菌光敏色素在2.1纳米分辨率的室温晶体结构。该结构是通过在X射线自由电子激光器下从微晶中连续飞秒X射线晶体学获得的。我们发现整体良好的协议相比,在1.35毫米的分辨率来自传统的晶体学在低温下,我们也在这里报告的晶体结构。发色团和蛋白质之间的硫醚键在同步加速器中受到位置模糊性的影响,但用SFX完全解决。该研究为利用时间分辨SFX研究光敏色素光循环的时间分辨结构铺平了道路。
Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
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