Conversion of red fluorescent protein into a bright blue probe.
Conversion of red fluorescent protein into a bright blue probe.
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DOI：
10.1016/j.chembiol.2008.08.006
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发表时间：
2008-10-20
影响因子：
--
通讯作者：
Verkhusha VV
中科院分区：
文献类型：
--
作者：
Subach OM;Gundorov IS;Yoshimura M;Subach FV;Zhang J;Grüenwald D;Souslova EA;Chudakov DM;Verkhusha VV
We utilized a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a novel rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic background such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which were converted into blue probes. Further improvement of a blue variant of TagRFP using random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by substantially higher brightness, faster chromophore maturation and higher pH stability than blue fluorescence proteins with a histidine in the chromophore. Detailed biochemical and photochemical analysis indicates mTagBFP is the true monomeric protein tag for multicolor and lifetime imaging as well as the outstanding donor for green fluorescent proteins in FRET applications.
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