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Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity

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 supr...

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Autores principales: Pina, Ana S., Morgado, Leonor, Duncan, Krystyna L., 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., Ulijn, Rein V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8694286/
https://www.ncbi.nlm.nih.gov/pubmed/35059169
http://dx.doi.org/10.1039/d1sc04420f
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author Pina, Ana S.
Morgado, Leonor
Duncan, Krystyna L.
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.
Ulijn, Rein V.
author_facet Pina, Ana S.
Morgado, Leonor
Duncan, Krystyna L.
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.
Ulijn, Rein V.
author_sort Pina, Ana S.
collection PubMed
description 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 with K(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 model para-nitro phenol phosphate substrate, with P7 demonstrating conformation-dependent activity with a modest k(cat)/K(M) = 4 ± 0.3 × 10(−4) M(−1) s(−1).
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spelling pubmed-86942862022-01-19 Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity Pina, Ana S. Morgado, Leonor Duncan, Krystyna L. 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. Ulijn, Rein V. Chem Sci Chemistry 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 with K(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 model para-nitro phenol phosphate substrate, with P7 demonstrating conformation-dependent activity with a modest k(cat)/K(M) = 4 ± 0.3 × 10(−4) M(−1) s(−1). The Royal Society of Chemistry 2021-12-07 /pmc/articles/PMC8694286/ /pubmed/35059169 http://dx.doi.org/10.1039/d1sc04420f Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Pina, Ana S.
Morgado, Leonor
Duncan, Krystyna L.
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.
Ulijn, Rein V.
Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title_full Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title_fullStr Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title_full_unstemmed Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title_short Discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
title_sort discovery of phosphotyrosine-binding oligopeptides with supramolecular target selectivity
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8694286/
https://www.ncbi.nlm.nih.gov/pubmed/35059169
http://dx.doi.org/10.1039/d1sc04420f
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