Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa.
Infrared spectroscopy reveals multi-step multi-timescale photoactivation in the photoconvertible protein archetype dronpa.
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DOI：
10.1038/s41557-018-0073-0
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发表时间：
2018-08
期刊：
影响因子：
21.8
通讯作者：
Meech SR
中科院分区：
文献类型：
--
作者：
Laptenok SP;Gil AA;Hall CR;Lukacs A;Iuliano JN;Jones GA;Greetham GM;Donaldson P;Miyawaki A;Tonge PJ;Meech SR
光致变色荧光蛋白在超分辨率显微镜和光遗传学中起着关键作用。调制荧光的光驱动结构变化涉及反式到顺式异构化和质子转移。发色团和蛋白质动力学的机制，时间尺度和相对贡献目前还没有很好地理解。在这里，“关”到“开”状态切换dronpa的机制进行了研究，使用飞秒到毫秒的时间分辨红外光谱和同位素标记。发色团和蛋白质的动力学发生在多个时间尺度上，从皮秒到数百微秒。在激发反式发色团之后，在皮秒内形成基态初级产物。令人惊讶的是，中性顺式异构体的特征振动光谱仅在几十纳秒后出现。需要中性顺式发色团周围的蛋白质结构的进一步波动以形成促进最终质子转移反应的新中间体。这些数据说明了发色团动力学和蛋白质环境之间的相互作用，荧光蛋白光致变色。
Photochromic fluorescent proteins play key roles in super-resolution microscopy and optogenetics. The light driven structure change which modulates the fluorescence involves both trans to cis isomerization and proton transfer. The mechanism, timescale and relative contribution of chromophore and protein dynamics are presently not well understood. Here the mechanism of ‘off’ to ‘on’ state switching in dronpa is studied using femtosecond to millisecond time resolved infra-red spectroscopy and isotope labelling. Chromophore and protein dynamics are shown to occur on multiple timescales, from picoseconds to hundreds of microseconds. Following excitation of the trans chromophore a ground state primary product is formed in picoseconds. Surprisingly the characteristic vibrational spectrum of the neutral cis isomer appears only after several tens of nanoseconds. Further fluctuations in protein structure around the neutral cis chromophore are required to form a new intermediate which promotes the final proton transfer reaction. These data illustrate the interplay between chromophore dynamics and protein environment underlying fluorescent protein photochromism.
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影响因子：
7.8
作者：
Zhou, Xin X.;Lin, Michael Z.
通讯作者：
Lin, Michael Z.
影响因子：
12.4
作者：
Shcherbakova DM;Sengupta P;Lippincott-Schwartz J;Verkhusha VV
通讯作者：
Verkhusha VV
影响因子：
5.8
作者：
Snellenburg, Joris J.;Laptenok, Sergey P.;van Stokkum, Ivo H. M.
通讯作者：
van Stokkum, Ivo H. M.
影响因子：
3.4
作者：
Ando, Ryoko;Flors, Cristina;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
DOI：
10.1093/protein/8.2.127
发表时间：
1995-02-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
WALLACE, AC;LASKOWSKI, RA;THORNTON, JM
通讯作者：
THORNTON, JM