Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing

Nat Commun. 2020 May 8;11(1):2284. doi: 10.1038/s41467-020-16208-6.


Manipulation of proteins by chemical modification is a powerful way to decipher their function. However, most ribosome-dependent and semi-synthetic methods have limitations in the number and type of modifications that can be introduced, especially in live cells. Here, we present an approach to incorporate single or multiple post-translational modifications or non-canonical amino acids into proteins expressed in eukaryotic cells. We insert synthetic peptides into GFP, NaV1.5 and P2X2 receptors via tandem protein trans-splicing using two orthogonal split intein pairs and validate our approach by investigating protein function. We anticipate the approach will overcome some drawbacks of existing protein enigineering methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Peptides / chemistry
  • Peptides / metabolism*
  • Protein Biosynthesis
  • Protein Domains
  • Protein Splicing*
  • Recombinant Proteins / metabolism
  • Trans-Splicing*
  • Xenopus laevis


  • Peptides
  • Recombinant Proteins
  • Green Fluorescent Proteins