Through the incorporation of the metal-chelating amino acid pyTyr into green fluorescent protein (GFP) from jellyfish, photoinduced electron transfer (ET) from the GFP chromophore to a bound Cu(II) ion was shown to occur within one nanosecond in a distance dependent manner. The crystal structure of GFP with pyTyr at a specific position shows the structural basis for the nanomolar binding affinity of pyTyr to Cu(II) ions.

The electron transfer (ET) involves many important biochemical processes in vivo, including photosynthesis. How to transform biological components to achieve efficient and controllable light-induced charge separation is an important problem and a major bottleneck means of synthetic biology to produce renewable energy. Meanwhile, the current experimental measurement of the electron transfer protein are usually rely connection probes to conduct a study on the protein itself contains a residue of histidine or cysteine, so that the method can only be used to study a small soluble protein, limits its application. Light-induced electron transfer (PET) The lead fluorescence quenching is a powerful tool used to explore the dynamic conformational changes of biological macromolecules. However, due to the limitations of current technology, only as an electron donor with a tyrosine or tryptophan.

The study will have a means of the metal chelating ability of non-natural amino acid via codon extended sentinel inserted into the green fluorescent protein (GFP), for the first time to achieve a rapid light-induced electron transfer between the fluorescent protein luminescent center to the copper ion, and measuring electron transfer occurs in a 1 nanosecond (near the center of the light velocity). The crystal structure reveals 3 - pyrazolyl tyrosine on the binding capacity of the copper ions have a high strength. These new methods of The protein dynamic conformational change research provides new means to provide new ideas for the use of synthetic biology means the production of renewable energy, provides a new tool for the design of metalloproteins. The paper also proposed jellyfish green fluorescent protein may be a new point of view of the jellyfish photoreceptors.

Princeton University, USA famous biophysical chemist Prof. Haw Yang council, said: "I believe that the non-natural amino acid coding technology researchers biophysics - the field of protein research will provide a very valuable tool in this article, is to promote one of the development in the field of research, because the use of copper as a quencher, can greatly expand the toolkit based on distance measurements. "

American University of Massachusetts famous bio-inorganic chemist Professor GUO Maolin, council said: "Understanding the biological electron transfer mechanism has been proved to be a challenging and complex scientific problems. Institute of Biophysics, Chinese Academy of Sciences, Dr. Wang and his colleagues developed a new strategy by the metal-chelating non-natural amino acids and encoded into proteins to study this complex issue. their success chelated Cu (II) groups in the green fluorescent protein (GFP) surface coding, 405nm light-induced light-induced electron transfer (PET) from protein chromophore of Cu (II) occurs rapidly, reducing the copper (I) and manufactured in a nanosecond. gene encoding strategy wonderful is that real-time monitoring to provide an opportunity for electron transfer proteins in living cells, which will be even more exciting! "

The Peking University famous biophysical chemist Professor Gao Yiqin council said: "light-induced electron transfer study of protein dynamics is a very useful tool, but its application is generally limited to a relatively simple system this work fine metal ion chelation together amino acids into proteins, which provide a new strategy for protein dynamics studies, this method is likely to significantly improve the applicability of PET in the study of protein dynamics. "


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