The Development of a Fluorescence-Based Competitive Assay Enabled the Discovery of Dimeric Cyclic Peptide Modulators of Ubiquitin Chains

Ganga B. Vamisetti, Roman Meledin, Mickal Nawatha, Hiroaki Suga, Ashraf Brik

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Development of modulators targeting specific interactions of ubiquitin-based conjugates with their partners is a formidable task since it requires a suitable screening assay and homogeneous ubiquitin conjugates. We developed a novel high-throughput strategy for screening ligands for Lys48-linked tetraubiquitin chain in a relatively simple, fast, and affordable manner. This approach combined with a state-of-the-art toolbox of chemical protein synthesis and a specially optimized Cys deprotection protocol enabled us to design highly potent, Lys48-linked tetraubiquitin chain selective “next generation” dimeric peptide modulators. The dimeric peptide exhibited cancer cell permeability and induced cell death with higher efficiency compared to its monocyclic analogue. These features make our dimeric peptide a promising candidate for further studies using in vivo models. Our assay can be adopted for other various ubiquitin chains in their free or anchored forms as well as conjugates for Ub-like modifiers.

Original languageEnglish
Pages (from-to)7018-7023
Number of pages6
JournalAngewandte Chemie - International Edition
Volume60
Issue number13
DOIs
StatePublished - 22 Mar 2021
Externally publishedYes

Keywords

  • cyclic peptides
  • dimeric peptides
  • fluorescence-based assay
  • high-throughput screening
  • peptide engineering

ASJC Scopus subject areas

  • Catalysis
  • General Chemistry

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