Verkhusha Vladislav V, Chudakov Dmitry M, Gurskaya Nadya G, Lukyanov Sergey, Lukyanov Konstantin A
Department of Pharmacology, University of Colorado Health Sciences Center, Denver, CO 80262, USA.
Chem Biol. 2004 Jun;11(6):845-54. doi: 10.1016/j.chembiol.2004.04.007.
The mechanism of the chromophore maturation in members of the green fluorescent protein (GFP) family such as DsRed and other red fluorescent and chromoproteins was analyzed. The analysis indicates that the red chromophore results from a chemical transformation of the protonated form of the GFP-like chromophore, not from the anionic form, which appears to be a dead-end product. The data suggest a rational strategy to achieve the complete red chromophore maturation utilizing substitutions to favor the formation of the neutral phenol in GFP-like chromophore. Our approach to detect the neutral chromophore form expands the application of fluorescent timer proteins to faster promoter activities and more spectrally distinguishable fluorescent colors. Light sensitivity found in the DsRed neutral form, resulting in its instant transformation to the mature red chromophore, could be exploited to accelerate the fluorescence acquisition.
对绿色荧光蛋白（GFP）家族成员如DsRed以及其他红色荧光蛋白和生色蛋白中生色团成熟的机制进行了分析。分析表明，红色生色团源自类GFP生色团质子化形式的化学转化，而非阴离子形式，阴离子形式似乎是一种终产物。数据表明了一种合理策略，即利用取代作用促进类GFP生色团中中性酚的形成，以实现完全的红色生色团成熟。我们检测中性生色团形式的方法将荧光计时蛋白的应用扩展到更快的启动子活性和更多光谱可区分的荧光颜色。在DsRed中性形式中发现的光敏感性导致其立即转化为成熟的红色生色团，可利用这一点加速荧光采集。
