Rational design of true monomeric and bright photoactivatable fluorescent proteins |
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Authors: | Zhang Mingshu Chang Hao Zhang Yongdeng Yu Junwei Wu Lijie Ji Wei Chen Juanjuan Liu Bei Lu Jingze Liu Yingfang Zhang Junlong Xu Pingyong Xu Tao |
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Affiliation: | National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China. |
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Abstract: | 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. |
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