Residue-specific incorporation of noncanonical amino acids for protein engineering

Mark B. van Eldijk, Jan C.M. van Hest

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

5 Citations (Scopus)


The incorporation of noncanonical amino acids has given protein chemists access to an expanded repertoire of amino acids. This methodology has significantly broadened the scope of protein engineering allowing introduction of amino acids with non-native functionalities, such as bioorthogonal reactive handles (azides and alkynes) and hydrophobic fluorinated side chains. Here, we describe the efficient residue-specific replacement of methionine by azidonorleucine in an engineered green fluorescent protein using a bacterial expression system to introduce a single reactive site for the strain-promoted azide-alkyne cycloaddition.

Original languageEnglish
Title of host publicationNoncanonical amino acids
Subtitle of host publicationmethods and protocols
EditorsE.A. Lemke
Place of PublicationNew York
PublisherHumana Press
Number of pages9
ISBN (Electronic) 978-1-4939-7574-7
ISBN (Print)978-1-4939-7573-0
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Aminoacyl-tRNA synthetase
  • Azidonorleucine
  • Bioconjugation
  • Medium shift
  • Noncanonical amino acids
  • Residue-specific incorporation
  • Strain-promoted azide-alkyne cycloaddition
  • Unnatural amino acids
  • Amino Acid Sequence
  • Gene Expression
  • Protein Biosynthesis
  • Amino Acids/chemistry
  • Azides/chemistry
  • Models, Molecular
  • Amino Acyl-tRNA Synthetases/chemistry
  • Norleucine/analogs & derivatives
  • Chromatography, Affinity
  • Proteins/chemistry
  • Protein Engineering
  • Protein Conformation
  • Genes, Reporter


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