earticle

영어

For the past decades, in vivo incorporation of Unnatural into recombinant protein expanded the application of protein with novel functions. Genetic code engineering (Codon reassignment) is one such methodology to design proteins with novel functional and biophysical properties. It is a recent developing method which engineer proteins with additional chemical moiety through incorporation of unnatural amino acids and helps in expanding the proteome of cell. To date, several groups successfully incorporated more than 100 unnatural amino acids into recombinant proteins through in vivo and in vitro approaches. Although, novel functional proteins were developed through this approach, we are the first to demonstrate the development of copper sensing fluorescent protein through unnatural amino acid mutagenesis. In this study we show the incorporation of unnatural amino acid, L-DOPA into Green fluorescent protein provides the protein with highly sensitive copper binding property. The Copper binding property of the engineered protein was characterized by the combination of fluorescent spectroscopy, absorption spectroscopy and circular dichroism. Overall this study demonstrates that the development of selective, sensitive and robust fluorescent based metal biosensor using unnatural amino acid mutagenesis.