title: Development of a Multifunctional Benzophenone Linker for Peptide Stapling and Photoaffinity Labelling.
abstract: 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.
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Yuteng
authors.initials: Y
authors.lastname: Wu
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Lasse B
authors.initials: LB
authors.lastname: Olsen
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Yu Heng
authors.initials: YH
authors.lastname: Lau
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Claus Hatt
authors.initials: CH
authors.lastname: Jensen
authors.affiliation: Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK.
authors.forename: Maxim
authors.initials: M
authors.lastname: Rossmann
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Ysobel R
authors.initials: YR
authors.lastname: Baker
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Hannah F
authors.initials: HF
authors.lastname: Sore
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK.
authors.forename: Súil
authors.initials: S
authors.lastname: Collins
authors.affiliation: University Chemical Laboratory, University of Cambridge, Lensfield Road, Cambridge, CB2 1EW, UK. spring@ch.cam.ac.uk.
authors.forename: David R
authors.initials: DR
authors.lastname: Spring
mesh_terms: D001577:Benzophenones / Q000138:chemical synthesis / Q000737:chemistry*; D057930:Click Chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D008024:Ligands; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry*; D019920:Photoaffinity Labels; D051736:Proto-Oncogene Proteins c-mdm2 / Q000737:chemistry*
chemical_list: D001577:Benzophenones; D003432:Cross-Linking Reagents; D008024:Ligands; D010455:Peptides; D019920:Photoaffinity Labels; C047723:benzophenone; D051736:Proto-Oncogene Proteins c-mdm2
keywords: MDM2; click chemistry; linker; photoaffinity labeling; stapled peptide