Genetic Incorporation of unnatural amino acids as electron acceptors

来源 :The 9th Asian Biophysics Association Symposium (ABA2015)(第九届 | 被引量 : 0次 | 上传用户:fh2029
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Electron transfer (ET) is widely used for driving the processes that underlie the chemistry of life.However, our abilities to probe electron transfer mechanisms in proteins and design redox enzymes are limited, due to the lack of methods to site-specifically insert electron acceptors into proteins in vivo.Here we describe the synthesis and genetic incorporation of 3-nitrophenylalanine (NO2Phe) and 4-fluoro-3-nitrophenylalanine (FNO2Phe), which are efficient electron acceptors with similar reduction potentials to NADH, the most important biological reductant.By genetic incorporation of NO2Phe and FNO2Phe into green fluorescent protein (GFP) and the use of femtosecond transient absorption measurement, we show that photo-induced electron transfer (PET) from the GFP chromophore to FNO2Phe occurs very fast (within 11 picoseconds), and is distance-dependent.These genetically encoded, low-reduction potential unnatural amino acids (UAA) can significantly improve our ability to investigate electron transfer mechanisms in complex reductases (such as hydrogenase and photosystem I), and facilitate the design of miniature proteins which mimic their functions.
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