Development of a multifunctional benzophenone linker for peptide stapling and photoaffinity labeling
Olsen, Lasse B
Lau, Yu Heng
Jensen, Claus Hatt
Baker, Ysobel R
MetadataShow full item record
Wu, Y., Olsen, L. B., Lau, Y. H., Jensen, C. H., Rossmann, M., Baker, Y. R., Sore, H., et al. (2016). Development of a multifunctional benzophenone linker for peptide stapling and photoaffinity labeling. ChemBioChem, 17 689-692. https://doi.org/10.1002/cbic.201500648
Photoaffinity labelling is a useful method for studying how proteins interact with ligands and biomolecules, and can help identify and characterise new targets for the development of new therapeutics. We present the design and synthesis of a novel multifunctional benzophenone linker, which serves as both a photocrosslinking motif and a peptide stapling reagent. Using a double-click stapling methodology, we attach the benzophenone to the peptide via the staple linker, rather than modifying the peptide sequence with a photocrosslinking amino acid. Applied to a p53-derived peptide, the resulting photoreactive stapled peptide is able to preferentially crosslink with MDM2 in the presence of competing protein. This multifunctional linker also features an extra alkyne handle for downstream applications such as pull-down assays, and can be used to investigate the target selectivity of stapled peptides.
photoaffinity labeling, stapled peptide, linker, click chemistry, MDM2
This work was supported by the EPSRC, BBSRC, MRC, Wellcome Trust and ERC (FP7/2007-2013; 279337/DOS). We thank Dr. Clemens Mayer for access to the UV crosslinker (University Chemical Laboratory, University of Cambridge), Weiyan Chen and Fran Kundel (University Chemical Laboratory, University of Cambridge) for assistance with the Typhoon imager and Dr. Laura Itzhaki and Wenshu Xu (Department of Pharmacology, University of Cambridge) for assistance with SDS-PAGE.
European Research Council (279337)
Embargo Lift Date
External DOI: https://doi.org/10.1002/cbic.201500648
This record's URL: https://www.repository.cam.ac.uk/handle/1810/254078