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Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions

Protein–peptide interactions are an essential player in cellular processes and, thus, of great interest as potential therapeutic agents. However, identifying the protein’s interacting surface has been shown to be a challenging task. Here, we present a methodology for protein–peptide interaction iden...

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Detalles Bibliográficos
Autores principales: Debeljak, Jerneja, Korošec, Peter, Šelb, Julij, Rijavec, Matija, Košnik, Mitja, Lunder, Mojca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953301/
https://www.ncbi.nlm.nih.gov/pubmed/36830679
http://dx.doi.org/10.3390/biom13020310
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author Debeljak, Jerneja
Korošec, Peter
Šelb, Julij
Rijavec, Matija
Košnik, Mitja
Lunder, Mojca
author_facet Debeljak, Jerneja
Korošec, Peter
Šelb, Julij
Rijavec, Matija
Košnik, Mitja
Lunder, Mojca
author_sort Debeljak, Jerneja
collection PubMed
description Protein–peptide interactions are an essential player in cellular processes and, thus, of great interest as potential therapeutic agents. However, identifying the protein’s interacting surface has been shown to be a challenging task. Here, we present a methodology for protein–peptide interaction identification, implementing phage panning, next-generation sequencing and bioinformatic analysis. One of the uses of this methodology is identification of allergen epitopes, especially suitable for globular inhaled and venom allergens, where their binding capability is determined by the allergen’s conformation, meaning their interaction cannot be properly studied when denatured. A Ph.D. commercial system based on the M13 phage vector was used for the panning process. Utilization of various bioinformatic tools, such as PuLSE, SAROTUP, MEME, Hammock and Pepitope, allowed us to evaluate a large amount of obtained data. Using the described methodology, we identified three peptide clusters representing potential epitopes on the major wasp venom allergen Ves v 5.
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spelling pubmed-99533012023-02-25 Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions Debeljak, Jerneja Korošec, Peter Šelb, Julij Rijavec, Matija Košnik, Mitja Lunder, Mojca Biomolecules Article Protein–peptide interactions are an essential player in cellular processes and, thus, of great interest as potential therapeutic agents. However, identifying the protein’s interacting surface has been shown to be a challenging task. Here, we present a methodology for protein–peptide interaction identification, implementing phage panning, next-generation sequencing and bioinformatic analysis. One of the uses of this methodology is identification of allergen epitopes, especially suitable for globular inhaled and venom allergens, where their binding capability is determined by the allergen’s conformation, meaning their interaction cannot be properly studied when denatured. A Ph.D. commercial system based on the M13 phage vector was used for the panning process. Utilization of various bioinformatic tools, such as PuLSE, SAROTUP, MEME, Hammock and Pepitope, allowed us to evaluate a large amount of obtained data. Using the described methodology, we identified three peptide clusters representing potential epitopes on the major wasp venom allergen Ves v 5. MDPI 2023-02-07 /pmc/articles/PMC9953301/ /pubmed/36830679 http://dx.doi.org/10.3390/biom13020310 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Debeljak, Jerneja
Korošec, Peter
Šelb, Julij
Rijavec, Matija
Košnik, Mitja
Lunder, Mojca
Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title_full Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title_fullStr Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title_full_unstemmed Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title_short Combination of Experimental and Bioinformatic Approaches for Identification of Immunologically Relevant Protein–Peptide Interactions
title_sort combination of experimental and bioinformatic approaches for identification of immunologically relevant protein–peptide interactions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953301/
https://www.ncbi.nlm.nih.gov/pubmed/36830679
http://dx.doi.org/10.3390/biom13020310
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