Identification of photocrosslinking peptide ligands by mRNA display
Abstract Photoaffinity labelling is a promising method for studying protein-ligand interactions. However, obtaining a specific, efficient crosslinker can require significant optimisation. We report a modified mRNA display strategy, photocrosslinking-RaPID (XL-RaPID), and exploit its ability to accel...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-05-01
|
| Series: | Communications Chemistry |
| Online Access: | https://doi.org/10.1038/s42004-023-00898-2 |
| _version_ | 1797778996066254848 |
|---|---|
| author | Yuteng Wu M. Teresa Bertran Dhira Joshi Sarah L. Maslen Catherine Hurd Louise J. Walport |
| author_facet | Yuteng Wu M. Teresa Bertran Dhira Joshi Sarah L. Maslen Catherine Hurd Louise J. Walport |
| author_sort | Yuteng Wu |
| collection | DOAJ |
| description | Abstract Photoaffinity labelling is a promising method for studying protein-ligand interactions. However, obtaining a specific, efficient crosslinker can require significant optimisation. We report a modified mRNA display strategy, photocrosslinking-RaPID (XL-RaPID), and exploit its ability to accelerate the discovery of cyclic peptides that photocrosslink to a target of interest. As a proof of concept, we generated a benzophenone-containing library and applied XL-RaPID screening against a model target, the second bromodomain of BRD3. This crosslinking screening gave two optimal candidates that selectively labelled the target protein in cell lysate. Overall, this work introduces direct photocrosslinking screening as a versatile technique for identifying covalent peptide ligands from mRNA display libraries incorporating reactive warheads. |
| first_indexed | 2024-03-12T23:25:27Z |
| format | Article |
| id | doaj.art-ac80405cd43a480cbf4202f8da9ed517 |
| institution | Directory Open Access Journal |
| issn | 2399-3669 |
| language | English |
| last_indexed | 2024-03-12T23:25:27Z |
| publishDate | 2023-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Chemistry |
| spelling | doaj.art-ac80405cd43a480cbf4202f8da9ed5172023-07-16T11:11:37ZengNature PortfolioCommunications Chemistry2399-36692023-05-01611910.1038/s42004-023-00898-2Identification of photocrosslinking peptide ligands by mRNA displayYuteng Wu0M. Teresa Bertran1Dhira Joshi2Sarah L. Maslen3Catherine Hurd4Louise J. Walport5Protein-Protein Interaction Laboratory, The Francis Crick InstituteProtein-Protein Interaction Laboratory, The Francis Crick InstituteChemical Biology, The Francis Crick InstituteProteomics, The Francis Crick InstituteProtein-Protein Interaction Laboratory, The Francis Crick InstituteProtein-Protein Interaction Laboratory, The Francis Crick InstituteAbstract Photoaffinity labelling is a promising method for studying protein-ligand interactions. However, obtaining a specific, efficient crosslinker can require significant optimisation. We report a modified mRNA display strategy, photocrosslinking-RaPID (XL-RaPID), and exploit its ability to accelerate the discovery of cyclic peptides that photocrosslink to a target of interest. As a proof of concept, we generated a benzophenone-containing library and applied XL-RaPID screening against a model target, the second bromodomain of BRD3. This crosslinking screening gave two optimal candidates that selectively labelled the target protein in cell lysate. Overall, this work introduces direct photocrosslinking screening as a versatile technique for identifying covalent peptide ligands from mRNA display libraries incorporating reactive warheads.https://doi.org/10.1038/s42004-023-00898-2 |
| spellingShingle | Yuteng Wu M. Teresa Bertran Dhira Joshi Sarah L. Maslen Catherine Hurd Louise J. Walport Identification of photocrosslinking peptide ligands by mRNA display Communications Chemistry |
| title | Identification of photocrosslinking peptide ligands by mRNA display |
| title_full | Identification of photocrosslinking peptide ligands by mRNA display |
| title_fullStr | Identification of photocrosslinking peptide ligands by mRNA display |
| title_full_unstemmed | Identification of photocrosslinking peptide ligands by mRNA display |
| title_short | Identification of photocrosslinking peptide ligands by mRNA display |
| title_sort | identification of photocrosslinking peptide ligands by mrna display |
| url | https://doi.org/10.1038/s42004-023-00898-2 |
| work_keys_str_mv | AT yutengwu identificationofphotocrosslinkingpeptideligandsbymrnadisplay AT mteresabertran identificationofphotocrosslinkingpeptideligandsbymrnadisplay AT dhirajoshi identificationofphotocrosslinkingpeptideligandsbymrnadisplay AT sarahlmaslen identificationofphotocrosslinkingpeptideligandsbymrnadisplay AT catherinehurd identificationofphotocrosslinkingpeptideligandsbymrnadisplay AT louisejwalport identificationofphotocrosslinkingpeptideligandsbymrnadisplay |