Liquid-microjet photoelectron spectroscopy of the green fluorescent protein chromophore.
Liquid-microjet photoelectron spectroscopy of the green fluorescent protein chromophore.
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DOI：
10.1038/s41467-022-28155-5
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发表时间：
2022-01-26
影响因子：
16.6
通讯作者：
Fielding HH
中科院分区：
文献类型：
--
作者：
Tau O;Henley A;Boichenko AN;Kleshchina NN;Riley R;Wang B;Winning D;Lewin R;Parkin IP;Ward JM;Hailes HC;Bochenkova AV;Fielding HH
绿色荧光蛋白（GFP）是用于体内监测生物过程的最广泛使用的荧光蛋白，已知其经历光氧化反应。然而，最基本的属性支撑光氧化，电子脱离能量，仅被测量为在气相中的去质子化的GFP发色团。在这里，我们使用多光子紫外光电子能谱在液体微射流和高层次的量子化学计算，以确定电子脱离能量的GFP发色团在水溶液中。发现与气相相比，水性环境提高了约4 eV的脱离能量，类似于在其天然蛋白质环境中的发色团的计算。在大多数情况下，电子脱离被发现发生共振通过电子激发态的生色团，突出其在光诱导的电子转移过程中的凝聚相的重要性。我们的研究结果表明，在水性环境中的GFP发色团的光氧化性能将是类似的蛋白质。光活性蛋白质的电子结构是许多自然光诱导过程的基础。作者，使用紫外液体微喷射光电子能谱和量子化学计算，确定电子脱离能量的绿色荧光蛋白生色团在水溶液中，接近蛋白质的环境条件。
Green fluorescent protein (GFP), the most widely used fluorescent protein for in vivo monitoring of biological processes, is known to undergo photooxidation reactions. However, the most fundamental property underpinning photooxidation, the electron detachment energy, has only been measured for the deprotonated GFP chromophore in the gas phase. Here, we use multiphoton ultraviolet photoelectron spectroscopy in a liquid-microjet and high-level quantum chemistry calculations to determine the electron detachment energy of the GFP chromophore in aqueous solution. The aqueous environment is found to raise the detachment energy by around 4 eV compared to the gas phase, similar to calculations of the chromophore in its native protein environment. In most cases, electron detachment is found to occur resonantly through electronically excited states of the chromophore, highlighting their importance in photo-induced electron transfer processes in the condensed phase. Our results suggest that the photooxidation properties of the GFP chromophore in an aqueous environment will be similar to those in the protein. The electronic structures of photoactive proteins underlie many natural photoinduced processes. The authors, using UV liquid-microjet photoelectron spectroscopy and quantum chemistry calculations, determine electron detachment energies of the green fluorescent protein chromophore in aqueous solution, approaching conditions of the protein environment.
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DOI：
10.1126/science.1207339
发表时间：
2011-07-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Paige JS;Wu KY;Jaffrey SR
通讯作者：
Jaffrey SR
影响因子：
8.6
作者：
Kiefer, Hjalte V.;Pedersen, Henrik B.;Andersen, Lars H.
通讯作者：
Andersen, Lars H.
影响因子：
8.4
作者：
Bochenkova AV;Mooney CRS;Parkes MA;Woodhouse JL;Zhang L;Lewin R;Ward JM;Hailes HC;Andersen LH;Fielding HH
通讯作者：
Fielding HH
影响因子：
64.8
作者：
Dickson, RM;Cubitt, AB;Moerner, WE
通讯作者：
Moerner, WE
影响因子：
3.3
作者：
Mandal, D;Tahara, T;Meech, SR
通讯作者：
Meech, SR