title: Exploring the substrate range of wild-type aminoacyl-tRNA synthetases.
abstract: We tested the substrate range of four wild-type E. coli aminoacyl-tRNA synthetases (AARSs) with a library of nonstandard amino acids (nsAAs). Although these AARSs could discriminate efficiently against the other canonical amino acids, they were able to use many nsAAs as substrates. Our results also show that E. coli tryptophanyl-tRNA synthetase (TrpRS) and tyrosyl-tRNA synthetase have overlapping substrate ranges. In addition, we found that the nature of the anticodon sequence of tRNA(Trp) altered the nsAA substrate range of TrpRS; this implies that the sequence of the anticodon affects the TrpRS amino acid binding pocket. These results highlight again that inherent AARS polyspecificity will be a major challenge in the aim of incorporating multiple different amino acids site-specifically into proteins.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520 (USA).
authors.forename: Chenguang
authors.initials: C
authors.lastname: Fan
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520 (USA).
authors.forename: Joanne M L
authors.initials: JML
authors.lastname: Ho
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520 (USA).
authors.forename: Napon
authors.initials: N
authors.lastname: Chirathivat
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520 (USA).|Department of Chemistry, Yale University, New Haven, CT 06520 (USA).
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520 (USA).
authors.forename: Yane-Shih
authors.initials: YS
authors.lastname: Wang
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics* / Q000378:metabolism*; D000926:Anticodon / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D008968:Molecular Conformation; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D000926:Anticodon; D000604:Amino Acyl-tRNA Synthetases
keywords: aminoacyl-tRNA synthetases; natural nonsense tRNA suppressors; nonstandard amino acids; substrate specificity; synthetic biology; tRNA