title: Site-specific incorporation of a fluorescent terphenyl unnatural amino acid.
abstract: The site-specific incorporation of unnatural amino acids into proteins has a wide range of biological implications. Of particular interest is the incorporation of fluorescent probes as a mechanism to track protein function, transport, and folding. Thus, the development of a novel system for the incorporation of new fluorescent unnatural amino acids has significant utility. Specifically, we have elucidated an aminoacyl-tRNA synthetase capable of recognizing a terphenyl UAA derivative, and charging a cognate tRNA with this amino acid for protein incorporation. Moreover, we have successfully incorporated this fluorescent UAA into GFP at several key residues, demonstrating a novel means to modulate fluorescence within the protein.
authors.affiliation: Department of Chemistry, College of William & Mary, Williamsburg, VA 23187, USA.
authors.forename: Jessica S
authors.initials: JS
authors.lastname: Lampkowski
authors.affiliation: Department of Chemistry, College of William & Mary, Williamsburg, VA 23187, USA.
authors.forename: Diya M
authors.initials: DM
authors.lastname: Uthappa
authors.affiliation: Department of Chemistry, College of William & Mary, Williamsburg, VA 23187, USA.
authors.forename: Douglas D
authors.initials: DD
authors.lastname: Young
mesh_terms: D019943:Amino Acid Substitution; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism*; D001713:Biphenyl Compounds / Q000138:chemical synthesis* / Q000378:metabolism; D004926:Escherichia coli; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D017434:Protein Structure, Tertiary; D013730:Terphenyl Compounds / Q000138:chemical synthesis* / Q000378:metabolism
chemical_list: C000608573:4-biphenyl-l-phenylalanine; D001713:Biphenyl Compounds; D005456:Fluorescent Dyes; D013730:Terphenyl Compounds; D049452:Green Fluorescent Proteins; D010649:Phenylalanine; D000604:Amino Acyl-tRNA Synthetases
keywords: Aminoacyl-tRNA synthetase; Fluorescent sensors; Green fluorescent protein; Protein expression; Unnatural amino acids