Double Strain‐Promoted Macrocyclization for the Rapid Selection of Cell‐Active Stapled Peptides. (2nd November 2015)

• Record Type: Journal Article
• Authors: Lau, Yu Heng; Wu, Yuteng; Rossmann, Maxim; Tan, Ban Xiong; de Andrade, Peterson; Tan, Yaw Sing; Verma, Chandra; McKenzie, Grahame J.; Venkitaraman, Ashok R.; Hyvönen, Marko; Spring, David R.
• Is Part Of: Angewandte Chemie. Volume 127:Number 51(2015)
• Page Start: 15630
• Page End: 15633
• Publication Date: 2015-11-02
• Subjects: Bioorthogonale Chemie -- Klick‐Chemie -- Makrocyclen -- Peptide -- Verklammerte Peptide; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201508416
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: Peptide stapling is a method for designing macrocyclic alpha‐helical inhibitors of protein–protein interactions. However, obtaining a cell‐active inhibitor can require significant optimization. We report a novel stapling technique based on a double strain‐promoted azide–alkyne reaction, and exploit its biocompatibility to accelerate the discovery of cell‐active stapled peptides. As a proof of concept, MDM2‐binding peptides were stapled in parallel, directly in cell culture medium in 96‐well plates, and simultaneously evaluated in a p53 reporter assay. This in situ stapling/screening process gave an optimal candidate that showed improved proteolytic stability and nanomolar binding to MDM2 in subsequent biophysical assays. α‐Helicity was confirmed by a crystal structure of the MDM2‐peptide complex. This work introduces in situ stapling as a versatile biocompatible technique with many other potential high‐throughput biological applications.