Chemical modification of proteins by insertion of synthetic peptides using tandem protein trans-splicing.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 32385250.
- Also identified by DOI 10.1038/s41467-020-16208-6 and PMC identifier 7210297.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
Abstract
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, Na<sub>V</sub>1.5 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.
Medical subject headings
- Peptides
- Protein Splicing
- Trans-Splicing