posted on 2021-11-05, 17:36authored byPramod
M. Sabale, Mateusz Imiołek, Pierre Raia, Sofia Barluenga, Nicolas Winssinger
Stapled peptides
with an enforced α-helical conformation
have been shown to overcome major limitations in the development of
short peptides targeting protein–protein interactions (PPIs).
While the growing arsenal of methodologies to staple peptides facilitates
their preparation, stapling methodologies are not broadly embraced
in synthetic library screening. Herein, we report a strategy leveraged
on hybridization of short PNA–peptide conjugates wherein nucleobase
driven assembly facilitates ligation of peptide fragments and constrains
the peptide’s conformation into an α-helix. Using native
chemical ligation, we show that a mixture of peptide fragments can
be combinatorially ligated and used directly in affinity selection
against a target of interest. This approach was exemplified with a
focused library targeting the p-53/MDM2 interaction. One hundred peptides
were obtained in a one-pot ligation reaction, selected by affinity
against MDM2 immobilized on beads, and the best binders were identified
by mass spectrometry.