Grison, CM, Burslem, GM, Miles, JA orcid.org/0000-0001-8839-9201 et al. (6 more authors) (2017) Double quick, double click reversible peptide “stapling”. Chemical Science, 8 (7). pp. 5166-5171. ISSN 2041-6520
Abstract
The development of constrained peptides for inhibition of protein–protein interactions is an emerging strategy in chemical biology and drug discovery. This manuscript introduces a versatile, rapid and reversible approach to constrain peptides in a bioactive helical conformation using BID and RNase S peptides as models. Dibromomaleimide is used to constrain BID and RNase S peptide sequence variants bearing cysteine (Cys) or homocysteine (hCys) amino acids spaced at i and i + 4 positions by double substitution. The constraint can be readily removed by displacement of the maleimide using excess thiol. This new constraining methodology results in enhanced α-helical conformation (BID and RNase S peptide) as demonstrated by circular dichroism and molecular dynamics simulations, resistance to proteolysis (BID) as demonstrated by trypsin proteolysis experiments and retained or enhanced potency of inhibition for Bcl-2 family protein–protein interactions (BID), or greater capability to restore the hydrolytic activity of the RNAse S protein (RNase S peptide). Finally, use of a dibromomaleimide functionalized with an alkyne permits further divergent functionalization through alkyne–azide cycloaddition chemistry on the constrained peptide with fluorescein, oligoethylene glycol or biotin groups to facilitate biophysical and cellular analyses. Hence this methodology may extend the scope and accessibility of peptide stapling.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Royal Society of Chemistry 2017. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material. |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Organic Chemistry (Leeds) |
Funding Information: | Funder Grant number EU - European Union 632207 EU - European Union 240324 Wellcome Trust 094232/Z/10/Z Leverhulme Trust RPG-2013-65 |
Depositing User: | Symplectic Publications |
Date Deposited: | 03 Jul 2017 10:58 |
Last Modified: | 24 Jul 2020 14:13 |
Status: | Published |
Publisher: | Royal Society of Chemistry |
Identification Number: | 10.1039/C7SC01342F |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:118497 |