Common pathway for the red chromophore formation in fluorescent proteins and chromoproteins
Common pathway for the red chromophore formation in fluorescent proteins and chromoproteins
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DOI：
10.1016/j.chembiol.2004.04.007
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发表时间：
2004-06-01
影响因子：
--
通讯作者：
Lukyanov, KA
中科院分区：
文献类型：
--
作者：
Verkhusha, VV;Chudakov, DM;Lukyanov, KA
分析了DsRed等绿色荧光蛋白（GFP）家族成员发色团成熟的机制。分析表明，红色发色团是由GFP样发色团的质子化形式的化学转化产生的，而不是由阴离子形式产生的，阴离子形式似乎是死端产物。这些数据表明了一种合理的策略，以实现完全的红色发色团成熟，利用取代有利于形成的中性酚类GFP样发色团。我们的方法来检测中性发色团的形式扩大了应用程序的荧光计时器蛋白更快的启动子活动和更光谱可区分的荧光颜色。在DsRed中性形式中发现的光敏感性，导致其瞬间转化为成熟的红色发色团，可以用来加速荧光采集。
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.