A rationally designed pyrrolysyl-tRNA synthetase mutant N346A/C348S was used for the genetic incorporation of fluorinated phenylalanine analogues pentafluorophenylalanine, 2,4,5,6-tetrafluorophenylalanine, 3,4,5-trifluorophenylalanine, and 3,4-difluorophenylalanine into superfolder green fluorescent protein (sfGFP) in Escherichia coli. Given that the comparable size of fluorine to hydrogen atom and fluorine's unique nuclear and electronegative properties, the fluorinated noncanonical amino acids have been used as probes to investigate protein folding and pi-pi interaction. Specifically, by introducing phenylalanine analogues with mono- or multiple- fluorination into a protein, the protein stability was able to be manipulated. The insight of sfGFP stability in terms of pi stacking was studied by incorporating fluorophenylalanine at various aromatic amino acid sites.

Manipulating protein pi-pi interactions by site-specific incorporation of fluorinated phenylalanine analogs

244th National Meeting (2012)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.