Development of a Multifunctional Benzophenone Linker for Peptide Stapling and Photoaffinity Labelling.

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Wu, Yuteng 
Olsen, Lasse B 
Lau, Yu Heng 
Jensen, Claus Hatt 

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 that serves as both a photo-crosslinking motif and a peptide stapling reagent. Using double-click stapling, we attached the benzophenone to the peptide via the staple linker, rather than by modifying the peptide sequence with a photo-crosslinking amino acid. When applied to a p53-derived peptide, the resulting photoreactive stapled peptide was 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.

MDM2, click chemistry, linker, photoaffinity labeling, stapled peptide, Benzophenones, Click Chemistry, Cross-Linking Reagents, Ligands, Molecular Structure, Peptides, Photoaffinity Labels, Proto-Oncogene Proteins c-mdm2
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Engineering and Physical Sciences Research Council (EP/K039520/1)
European Research Council (279337)
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.