Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity

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Pina, Ana S. and Morgado, Leonor and Duncan, Krystyna L. and Carvalho, Sara and Carvalho, Henrique F. and Barbosa, Arménio J. M. and de P. Mariz, Beatriz and Moreira, Inês P. and Kalafatovic, Daniela and Morais Faustino, Bruno M. and Narang, Vishal and Wang, Tong and Pappas, Charalampos G. and Ferreira, Isabel and Roque, A. Cecília A. and Ulijn, Rein V. (2022) Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity. Chemical Science, 13 (1). pp. 210-217. ISSN 2041-6539 (https://doi.org/10.1039/d1sc04420f)

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Abstract

We demonstrate phage-display screening on self-assembled ligands that enables the identification of oligopeptides that selectively bind dynamic supramolecular targets over their unassembled counterparts. The concept is demonstrated through panning of a phage-display oligopeptide library against supramolecular tyrosine-phosphate ligands using 9-fluorenylmethoxycarbonyl-phenylalanine-tyrosine-phosphate (Fmoc-FpY) micellar aggregates as targets. The 14 selected peptides showed no sequence consensus but were enriched in cationic and proline residues. The lead peptide, KVYFSIPWRVPM-NH 2(P7) was found to bind to the Fmoc-FpY ligand exclusively in its self-assembled state withK D= 74 ± 3 μM. Circular dichroism, NMR and molecular dynamics simulations revealed that the peptide interacts with Fmoc-FpY through the KVYF terminus and this binding event disrupts the assembled structure. In absence of the target micellar aggregate, P7 was further found to dynamically alternate between multiple conformations, with a preferred hairpin-like conformation that was shown to contribute to supramolecular ligand binding. Three identified phages presented appreciable binding, and two showed to catalyze the hydrolysis of a modelpara-nitro phenol phosphate substrate, with P7 demonstrating conformation-dependent activity with a modestk cat/K M= 4 ± 0.3 × 10 −4M −1s −1

ORCID iDs

Pina, Ana S., Morgado, Leonor, Duncan, Krystyna L. ORCID logoORCID: https://orcid.org/0000-0002-0654-0871, Carvalho, Sara, Carvalho, Henrique F., Barbosa, Arménio J. M., de P. Mariz, Beatriz, Moreira, Inês P., Kalafatovic, Daniela, Morais Faustino, Bruno M., Narang, Vishal, Wang, Tong, Pappas, Charalampos G., Ferreira, Isabel, Roque, A. Cecília A. and Ulijn, Rein V. ORCID logoORCID: https://orcid.org/0000-0001-7974-3779;
  • Item type:Article
    ID code:79121
    Dates:
    Date
    Event
    7 January 2022
    Published
    7 December 2021
    Published Online
    5 December 2021
    Accepted
    Subjects:Science > Chemistry
    Department:Faculty of Science > Pure and Applied Chemistry
    Depositing user: Pure Administrator
    Date deposited:13 Jan 2022 16:10
    Last modified:30 Nov 2024 14:18
    URI:https://strathprints.strath.ac.uk/id/eprint/79121
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