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《中国化学会-生物物理化学专业委员会第四届全国生物物理化学会议论文集》2016年
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Genetic Incorporation of Unnatural Amino Acid as Protein Electron Transfer Probes

刘晓红  
【摘要】:Electron transfer(ET) reactions govern many important biochemical processes, including oxygen binding, photosynthesis, respiration, and detoxification. Efficient photo-induced electron transfer(PET) typically requires close proximity of the donor and acceptor. The PET method nicely complements FRET studies for the investigation of small-scale conformation changes in biopolymers. By using of genetic incorporation of unnatural amino acids(UAA), several UAAs can be site-specifically incorporated into fluorescent proteins. We showed that PET from the chromophore to UAA or their metal complex can occur rapidly, suggesting that the fluorescent proteins may be optimized for the ET reaction. By substituting Tyr of the chromophore in diverse FPs with Hq Ala, we show for the first time UAA incorporation result in significantly red-shifted excitation and emission maxima. Such red-shifted GFP variants are extremely useful for tracking protein localization in cells, deep-tissue imaging, and the construction of fluorescence resonance energy transfer(FRET) sensors for a host of signaling molecules and post-translational modifications. These new genetically encoded amino acids could also aid the rational design of metalloproteins, as well as protein NMR spectroscopy using paramagnetic ions.
【作者单位】:中国科学院生物物理研究所
【分类号】:Q51

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