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Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition
Understanding the relationship between protein sequence and molecular recognition selectivity remains a major challenge. The antibody fragment scFv1F4 recognizes with sub nM affinity a decapeptide (sequence (6)TAMFQDPQER(15)) derived from the N-terminal end of human papilloma virus E6 oncoprotein. U...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4667898/ https://www.ncbi.nlm.nih.gov/pubmed/26629896 http://dx.doi.org/10.1371/journal.pone.0143374 |
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author | Nominé, Yves Choulier, Laurence Travé, Gilles Vernet, Thierry Altschuh, Danièle |
author_facet | Nominé, Yves Choulier, Laurence Travé, Gilles Vernet, Thierry Altschuh, Danièle |
author_sort | Nominé, Yves |
collection | PubMed |
description | Understanding the relationship between protein sequence and molecular recognition selectivity remains a major challenge. The antibody fragment scFv1F4 recognizes with sub nM affinity a decapeptide (sequence (6)TAMFQDPQER(15)) derived from the N-terminal end of human papilloma virus E6 oncoprotein. Using this decapeptide as antigen, we had previously shown that only the wild type amino-acid or conservative replacements were allowed at positions 9 to 12 and 15 of the peptide, indicating a strong binding selectivity. Nevertheless phenylalanine (F) was equally well tolerated as the wild type glutamine (Q) at position 13, while all other amino acids led to weaker scFv binding. The interfaces of complexes involving either Q or F are expected to diverge, due to the different physico-chemistry of these residues. This would imply that high-affinity binding can be achieved through distinct interfacial geometries. In order to investigate this point, we disrupted the scFv–peptide interface by modifying one or several peptide positions. We then analyzed the effect on binding of amino acid changes at the remaining positions, an altered susceptibility being indicative of an altered role in complex formation. The 23 starting variants analyzed contained replacements whose effects on scFv1F4 binding ranged from minor to drastic. A permutation analysis (effect of replacing each peptide position by all other amino acids except cysteine) was carried out on the 23 variants using the PEPperCHIP® Platform technology. A comparison of their permutation patterns with that of the wild type peptide indicated that starting replacements at position 11, 12 or 13 modified the tolerance to amino-acid changes at the other two positions. The interdependence between the three positions was confirmed by SPR (Biacore® technology). Our data demonstrate that binding selectivity does not preclude the existence of alternative high-affinity recognition modes. |
format | Online Article Text |
id | pubmed-4667898 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-46678982015-12-10 Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition Nominé, Yves Choulier, Laurence Travé, Gilles Vernet, Thierry Altschuh, Danièle PLoS One Research Article Understanding the relationship between protein sequence and molecular recognition selectivity remains a major challenge. The antibody fragment scFv1F4 recognizes with sub nM affinity a decapeptide (sequence (6)TAMFQDPQER(15)) derived from the N-terminal end of human papilloma virus E6 oncoprotein. Using this decapeptide as antigen, we had previously shown that only the wild type amino-acid or conservative replacements were allowed at positions 9 to 12 and 15 of the peptide, indicating a strong binding selectivity. Nevertheless phenylalanine (F) was equally well tolerated as the wild type glutamine (Q) at position 13, while all other amino acids led to weaker scFv binding. The interfaces of complexes involving either Q or F are expected to diverge, due to the different physico-chemistry of these residues. This would imply that high-affinity binding can be achieved through distinct interfacial geometries. In order to investigate this point, we disrupted the scFv–peptide interface by modifying one or several peptide positions. We then analyzed the effect on binding of amino acid changes at the remaining positions, an altered susceptibility being indicative of an altered role in complex formation. The 23 starting variants analyzed contained replacements whose effects on scFv1F4 binding ranged from minor to drastic. A permutation analysis (effect of replacing each peptide position by all other amino acids except cysteine) was carried out on the 23 variants using the PEPperCHIP® Platform technology. A comparison of their permutation patterns with that of the wild type peptide indicated that starting replacements at position 11, 12 or 13 modified the tolerance to amino-acid changes at the other two positions. The interdependence between the three positions was confirmed by SPR (Biacore® technology). Our data demonstrate that binding selectivity does not preclude the existence of alternative high-affinity recognition modes. Public Library of Science 2015-12-02 /pmc/articles/PMC4667898/ /pubmed/26629896 http://dx.doi.org/10.1371/journal.pone.0143374 Text en © 2015 Nominé et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Nominé, Yves Choulier, Laurence Travé, Gilles Vernet, Thierry Altschuh, Danièle Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title | Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title_full | Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title_fullStr | Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title_full_unstemmed | Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title_short | Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition |
title_sort | antibody binding selectivity: alternative sets of antigen residues entail high-affinity recognition |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4667898/ https://www.ncbi.nlm.nih.gov/pubmed/26629896 http://dx.doi.org/10.1371/journal.pone.0143374 |
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