Rational design of true monomeric and bright photoactivatable fluorescent proteins
Rational design of true monomeric and bright photoactivatable fluorescent proteins
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真正的单体和明亮的光激活荧光蛋白的合理设计
DOI：
10.1038/nmeth.2021
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发表时间：
2012-07-01
期刊：
影响因子：
48
通讯作者：
Xu, Tao
中科院分区：
文献类型：
--
作者：
Zhang, Mingshu;Chang, Hao;Xu, Tao
单体（m）Eos 2是一种工程化的可光活化的荧光蛋白，广泛用于超分辨率显微镜。我们发现，mEos2在高浓度下形成低聚物，并在标记膜蛋白时形成聚集体，限制了其作为融合伴侣的应用。我们解决了四聚体mEos 2的晶体结构，并合理设计了改进的版本mEos 3.1和mEos 3.2，它们是真正的单体，更亮，成熟更快，并表现出更高的光子预算和标记密度。
Monomeric (m)Eos2 is an engineered photoactivatable fluorescent protein widely used for super-resolution microscopy. We show that mEos2 forms oligomers at high concentrations and forms aggregates when labeling membrane proteins, limiting its application as a fusion partner. We solved the crystal structure of tetrameric mEos2 and rationally designed improved versions, mEos3.1 and mEos3.2, that are truly monomeric, are brighter, mature faster and exhibit higher photon budget and label density.