Bozhanova Nina G, Baranov Mikhail S, Sarkisyan Karen S, Gritcenko Roman, Mineev Konstantin S, Golodukhina Svetlana V, Baleeva Nadezhda S, Lukyanov Konstantin A, Mishin Alexander S
Institute of Bioorganic Chemistry, Russian Academy of Sciences , Miklukho-Maklaya 16/10, 117997 Moscow, Russia.
Pirogov Russian National Research Medical University , Ostrovitianov 1, 117997 Moscow, Russia.
ACS Chem Biol. 2017 Jul 21;12(7):1867-1873. doi: 10.1021/acschembio.7b00337. Epub 2017 Jun 6.
Rapid development of new microscopy techniques exposed the need for genetically encoded fluorescent tags with special properties. Recent works demonstrated the potential of fluorescent proteins with tryptophan-based chromophores. We applied rational design and random mutagenesis to the monomeric red fluorescent protein FusionRed and found two groups of mutants carrying a tryptophan-based chromophore: with yellow (535 nm) or orange (565 nm) emission. On the basis of the properties of proteins, a model synthetic chromophore, and a computational modeling, we concluded that the presence of a ketone-containing chromophore in different isomeric forms can explain the observed yellow and orange phenotypes.
新显微镜技术的快速发展凸显了对具有特殊性质的基因编码荧光标签的需求。近期研究表明了基于色氨酸的发色团的荧光蛋白的潜力。我们对单体红色荧光蛋白FusionRed应用了理性设计和随机诱变，发现了两组携带基于色氨酸的发色团的突变体：发射黄色（535纳米）或橙色（565纳米）荧光。基于蛋白质性质、模型合成发色团和计算建模，我们得出结论，不同异构体形式的含酮发色团的存在可以解释观察到的黄色和橙色表型。
