Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins
Exploration of new chromophore structures leads to the identification of improved blue fluorescent proteins
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DOI：
10.1021/bi700199g
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发表时间：
2007-05-22
期刊：
影响因子：
2.9
通讯作者：
Campbell, Robert E.
中科院分区：
文献类型：
--
作者：
Ai, Hui-wang;Shaner, Nathan C.;Campbell, Robert E.
水母绿色荧光蛋白（GFP）的变体称为蓝色荧光蛋白（BFP），最初是通过用组氨酸取代发色团前体序列中的酪氨酸而工程化的。在此，我们报告了BFP的改进版本沿着各种工程化荧光蛋白变体，其具有新颖且独特的发色团结构，所有发色团结构都具有蓝色荧光色调的性质。两个最有趣的新变体是GFP的一个版本，其中发色团不经历激发态质子转移，以及具有苯丙氨酸衍生发色团的mCherry版本。所有新的蓝色荧光蛋白质都已被严格评估其在活细胞荧光成像中的实用性。这些新的变异应该大大促进荧光成像的合法化蓝色荧光蛋白作为实用和强大的成员的荧光蛋白“工具包”。
The variant of Aequorea green fluorescent protein (GFP) known as blue fluorescent protein (BFP) was originally engineered by substituting histidine for tyrosine in the chromophore precursor sequence. Herein we report improved versions of BFP along with a variety of engineered fluorescent protein variants with novel and distinct chromophore structures that all share the property of a blue fluorescent hue. The two most intriguing of the new variants are a version of GFP in which the chromophore does not undergo excited-state proton transfer and a version of mCherry with a phenylalanine-derived chromophore. All of the new blue fluorescing proteins have been critically assessed for their utility in live cell fluorescent imaging. These new variants should greatly facilitate multicolor fluorescent imaging by legitimizing blue fluorescing proteins as practical and robust members of the fluorescent protein "toolkit".