A rationally designed pyrrolysyl-tRNA synthetase mutant with a broad substrate spectrum

J Am Chem Soc. 2012 Feb 15;134(6):2950-3. doi: 10.1021/ja211972x. Epub 2012 Feb 6.

Abstract

Together with tRNA(CUA)(Pyl), a rationally designed pyrrolysyl-tRNA synthetase mutant N346A/C348A has been successfully used for the genetic incorporation of a variety of phenylalanine derivatives with large para substituents into superfolder green fluorescent protein at an amber mutation site in Escherichia coli. This discovery greatly expands the genetically encoded noncanonical amino acid inventory and opens the gate for the genetic incorporation of other phenylalanine derivatives using engineered pyrrolysyl-tRNA synthetase-tRNA(CUA)(Pyl) pairs.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acyl-tRNA Synthetases / genetics*
  • Binding Sites
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / metabolism
  • Genetic Engineering / methods
  • Green Fluorescent Proteins / chemistry
  • Models, Chemical
  • Mutation*
  • Nucleic Acid Conformation
  • Peptides / chemistry
  • Phenylalanine / chemistry
  • Protein Engineering / methods*
  • Pyrroles / chemistry*
  • Substrate Specificity

Substances

  • Peptides
  • Pyrroles
  • Green Fluorescent Proteins
  • Phenylalanine
  • Amino Acyl-tRNA Synthetases