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Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries
Characterizing the binding selectivity landscape of interacting proteins is crucial both for elucidating the underlying mechanisms of their interaction and for developing selective inhibitors. However, current mapping methods are laborious and cannot provide a sufficiently comprehensive description...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158287/ https://www.ncbi.nlm.nih.gov/pubmed/30258049 http://dx.doi.org/10.1038/s41467-018-06403-x |
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author | Naftaly, Si Cohen, Itay Shahar, Anat Hockla, Alexandra Radisky, Evette S. Papo, Niv |
author_facet | Naftaly, Si Cohen, Itay Shahar, Anat Hockla, Alexandra Radisky, Evette S. Papo, Niv |
author_sort | Naftaly, Si |
collection | PubMed |
description | Characterizing the binding selectivity landscape of interacting proteins is crucial both for elucidating the underlying mechanisms of their interaction and for developing selective inhibitors. However, current mapping methods are laborious and cannot provide a sufficiently comprehensive description of the landscape. Here, we introduce a novel and efficient strategy for comprehensively mapping the binding landscape of proteins using a combination of experimental multi-target selective library screening and in silico next-generation sequencing analysis. We map the binding landscape of a non-selective trypsin inhibitor, the amyloid protein precursor inhibitor (APPI), to each of the four human serine proteases (kallikrein-6, mesotrypsin, and anionic and cationic trypsins). We then use this map to dissect and improve the affinity and selectivity of APPI variants toward each of the four proteases. Our strategy can be used as a platform for the development of a new generation of target-selective probes and therapeutic agents based on selective protein–protein interactions. |
format | Online Article Text |
id | pubmed-6158287 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-61582872018-10-01 Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries Naftaly, Si Cohen, Itay Shahar, Anat Hockla, Alexandra Radisky, Evette S. Papo, Niv Nat Commun Article Characterizing the binding selectivity landscape of interacting proteins is crucial both for elucidating the underlying mechanisms of their interaction and for developing selective inhibitors. However, current mapping methods are laborious and cannot provide a sufficiently comprehensive description of the landscape. Here, we introduce a novel and efficient strategy for comprehensively mapping the binding landscape of proteins using a combination of experimental multi-target selective library screening and in silico next-generation sequencing analysis. We map the binding landscape of a non-selective trypsin inhibitor, the amyloid protein precursor inhibitor (APPI), to each of the four human serine proteases (kallikrein-6, mesotrypsin, and anionic and cationic trypsins). We then use this map to dissect and improve the affinity and selectivity of APPI variants toward each of the four proteases. Our strategy can be used as a platform for the development of a new generation of target-selective probes and therapeutic agents based on selective protein–protein interactions. Nature Publishing Group UK 2018-09-26 /pmc/articles/PMC6158287/ /pubmed/30258049 http://dx.doi.org/10.1038/s41467-018-06403-x Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Naftaly, Si Cohen, Itay Shahar, Anat Hockla, Alexandra Radisky, Evette S. Papo, Niv Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title | Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title_full | Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title_fullStr | Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title_full_unstemmed | Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title_short | Mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
title_sort | mapping protein selectivity landscapes using multi-target selective screening and next-generation sequencing of combinatorial libraries |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158287/ https://www.ncbi.nlm.nih.gov/pubmed/30258049 http://dx.doi.org/10.1038/s41467-018-06403-x |
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