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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...
Autores principales: | , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Royal Society of Chemistry
2021
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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). |
format | Online Article Text |
id | pubmed-8694286 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
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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