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Double Strain-Promoted Macrocyclization for the Rapid Selection of Cell-Active Stapled Peptides.

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.
Source :
Angewandte Chemie International Edition; 12/14/2015, Vol. 54 Issue 51, p15410-15413, 4p, 1 Color Photograph, 2 Diagrams, 2 Graphs
Publication Year :
2015

Abstract

Peptide stapling is a method for designing macro-cyclic 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. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
14337851
Volume :
54
Issue :
51
Database :
Complementary Index
Journal :
Angewandte Chemie International Edition
Publication Type :
Academic Journal
Accession number :
111659329
Full Text :
https://doi.org/10.1002/anie.201508416