Observation of Cation Chromophore Photoisomerization of a Fluorescent Protein Using Millisecond Synchrotron Serial Crystallography and Infrared Vibrational and Visible Spectroscopy.
Observation of Cation Chromophore Photoisomerization of a Fluorescent Protein Using Millisecond Synchrotron Serial Crystallography and Infrared Vibrational and Visible Spectroscopy.
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DOI：
10.1021/acs.jpcb.2c06780
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发表时间：
2022-11-17
影响因子：
3.3
通讯作者：
van Thor, Jasper J.
中科院分区：
文献类型：
--
作者：
Baxter, James M.;Hutchison, Christopher D. M.;Maghlaoui, Karim;Cordon-Preciado, Violeta;Aller, Pierre;Butryn, Agata;Axford, Danny;Horrell, Sam;Owen, Robin L.;Storm, Selina L. S.;Devenish, Nicholas E.;van Thor, Jasper J.
可逆切换荧光蛋白（rsFP）的发色团经历反式和顺式形式的光异构化。与顺式/反式光异构化同时，rsFP通常在酚氧上质子化，导致吸收蓝移。合成的rsFP称为rsEospa，来自EosFP家族，在pH 8.4下显示与GFP样rsFP Dronpa相同的光谱行为，并且涉及非荧光中性和荧光阴离子发色团状态之间的光转换。毫秒时间分辨同步辐射系列晶体学的rsEospa在pH值8.4表明，光异构化是伴随着相同的三个残基中看到的Dronpa的重排。然而，在pH 5.5时，我们观察到OFF状态被鉴定为具有咪唑啉酮氮的额外质子化的阳离子发色团，所述咪唑啉酮氮与Glu 212羧酸酯侧链的新形成的氢键同时发生。FTIR光谱解析了阳离子物质在1713 cm-1处的特征上移羰基伸缩频率。此外，电子光谱将397 nm处的阳离子吸收带与387 nm处观察到的pH 8.4下的中性物质区分开。的阳离子发色团状态的光异构化的观察表明，圆锥形交叉的电子配置，其中先前荧光被提出是主要的衰减途径，为国家含有咪唑啉酮氮质子化。我们提出了完整的时间分辨室温X射线晶体学，FTIR，紫外/维斯分配和光转换建模的rsEospa。
The chromophores of reversibly switchable fluorescent proteins (rsFPs) undergo photoisomerization of both the trans and cis forms. Concurrent with cis/trans photoisomerisation, rsFPs typically become protonated on the phenolic oxygen resulting in a blue shift of the absorption. A synthetic rsFP referred to as rsEospa, derived from EosFP family, displays the same spectroscopic behavior as the GFP-like rsFP Dronpa at pH 8.4 and involves the photoconversion between nonfluorescent neutral and fluorescent anionic chromophore states. Millisecond time-resolved synchrotron serial crystallography of rsEospa at pH 8.4 shows that photoisomerization is accompanied by rearrangements of the same three residues as seen in Dronpa. However, at pH 5.5 we observe that the OFF state is identified as the cationic chromophore with additional protonation of the imidazolinone nitrogen which is concurrent with a newly formed hydrogen bond with the Glu212 carboxylate side chain. FTIR spectroscopy resolves the characteristic up-shifted carbonyl stretching frequency at 1713 cm–1 for the cationic species. Electronic spectroscopy furthermore distinguishes the cationic absorption band at 397 nm from the neutral species at pH 8.4 seen at 387 nm. The observation of photoisomerization of the cationic chromophore state demonstrates the conical intersection for the electronic configuration, where previously fluorescence was proposed to be the main decay route for states containing imidazolinone nitrogen protonation. We present the full time-resolved room-temperature X-ray crystallographic, FTIR, and UV/vis assignment and photoconversion modeling of rsEospa.
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影响因子：
21.8
作者：
Laptenok SP;Gil AA;Hall CR;Lukacs A;Iuliano JN;Jones GA;Greetham GM;Donaldson P;Miyawaki A;Tonge PJ;Meech SR
通讯作者：
Meech SR
DOI：
10.1073/pnas.0500489102
发表时间：
2005-07-05
影响因子：
11.1
作者：
Habuchi, S;Ando, R;Hofkens, J
通讯作者：
Hofkens, J
影响因子：
3.7
作者：
Arpino JA;Rizkallah PJ;Jones DD
通讯作者：
Jones DD
影响因子：
2.9
作者：
Elsliger, MA;Wachter, RM;Remington, SJ
通讯作者：
Remington, SJ
影响因子：
9.8
作者：
Lambert GG;Depernet H;Gotthard G;Schultz DT;Navizet I;Lambert T;Adams SR;Torreblanca-Zanca A;Chu M;Bindels DS;Levesque V;Nero Moffatt J;Salih A;Royant A;Shaner NC
通讯作者：
Shaner NC