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De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries
The α-helix is one of the most common protein surface recognition motifs found in nature, and its unique amide-cloaking properties also enable α-helical polypeptide motifs to exist in membranes. Together, these properties have inspired the development of α-helically constrained (Helicon) therapeutic...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9907135/ https://www.ncbi.nlm.nih.gov/pubmed/36534810 http://dx.doi.org/10.1073/pnas.2210435119 |
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author | Li, Kunhua Tokareva, Olena S. Thomson, Ty M. Wahl, Sebastian C. T. Travaline, Tara L. Ramirez, Jessica D. Choudary, Santosh K. Agarwal, Sorabh Walkup, Ward G. Olsen, Tivoli J. Brennan, Matthew J. Verdine, Gregory L. McGee, John H. |
author_facet | Li, Kunhua Tokareva, Olena S. Thomson, Ty M. Wahl, Sebastian C. T. Travaline, Tara L. Ramirez, Jessica D. Choudary, Santosh K. Agarwal, Sorabh Walkup, Ward G. Olsen, Tivoli J. Brennan, Matthew J. Verdine, Gregory L. McGee, John H. |
author_sort | Li, Kunhua |
collection | PubMed |
description | The α-helix is one of the most common protein surface recognition motifs found in nature, and its unique amide-cloaking properties also enable α-helical polypeptide motifs to exist in membranes. Together, these properties have inspired the development of α-helically constrained (Helicon) therapeutics that can enter cells and bind targets that have been considered “undruggable”, such as protein–protein interactions. To date, no general method for discovering α-helical binders to proteins has been reported, limiting Helicon drug discovery to only those proteins with previously characterized α-helix recognition sites, and restricting the starting chemical matter to those known α-helical binders. Here, we report a general and rapid screening method to empirically map the α-helix binding sites on a broad range of target proteins in parallel using large, unbiased Helicon phage display libraries and next-generation sequencing. We apply this method to screen six structurally diverse protein domains, only one of which had been previously reported to bind isolated α-helical peptides, discovering 20 families that collectively comprise several hundred individual Helicons. Analysis of 14 X-ray cocrystal structures reveals at least nine distinct α-helix recognition sites across these six proteins, and biochemical and biophysical studies show that these Helicons can block protein–protein interactions, inhibit enzymatic activity, induce conformational rearrangements, and cause protein dimerization. We anticipate that this method will prove broadly useful for the study of protein recognition and for the development of both biochemical tools and therapeutics for traditionally challenging protein targets. |
format | Online Article Text |
id | pubmed-9907135 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-99071352023-02-08 De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries Li, Kunhua Tokareva, Olena S. Thomson, Ty M. Wahl, Sebastian C. T. Travaline, Tara L. Ramirez, Jessica D. Choudary, Santosh K. Agarwal, Sorabh Walkup, Ward G. Olsen, Tivoli J. Brennan, Matthew J. Verdine, Gregory L. McGee, John H. Proc Natl Acad Sci U S A Biological Sciences The α-helix is one of the most common protein surface recognition motifs found in nature, and its unique amide-cloaking properties also enable α-helical polypeptide motifs to exist in membranes. Together, these properties have inspired the development of α-helically constrained (Helicon) therapeutics that can enter cells and bind targets that have been considered “undruggable”, such as protein–protein interactions. To date, no general method for discovering α-helical binders to proteins has been reported, limiting Helicon drug discovery to only those proteins with previously characterized α-helix recognition sites, and restricting the starting chemical matter to those known α-helical binders. Here, we report a general and rapid screening method to empirically map the α-helix binding sites on a broad range of target proteins in parallel using large, unbiased Helicon phage display libraries and next-generation sequencing. We apply this method to screen six structurally diverse protein domains, only one of which had been previously reported to bind isolated α-helical peptides, discovering 20 families that collectively comprise several hundred individual Helicons. Analysis of 14 X-ray cocrystal structures reveals at least nine distinct α-helix recognition sites across these six proteins, and biochemical and biophysical studies show that these Helicons can block protein–protein interactions, inhibit enzymatic activity, induce conformational rearrangements, and cause protein dimerization. We anticipate that this method will prove broadly useful for the study of protein recognition and for the development of both biochemical tools and therapeutics for traditionally challenging protein targets. National Academy of Sciences 2022-12-19 2022-12-27 /pmc/articles/PMC9907135/ /pubmed/36534810 http://dx.doi.org/10.1073/pnas.2210435119 Text en Copyright © 2022 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Biological Sciences Li, Kunhua Tokareva, Olena S. Thomson, Ty M. Wahl, Sebastian C. T. Travaline, Tara L. Ramirez, Jessica D. Choudary, Santosh K. Agarwal, Sorabh Walkup, Ward G. Olsen, Tivoli J. Brennan, Matthew J. Verdine, Gregory L. McGee, John H. De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title | De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title_full | De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title_fullStr | De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title_full_unstemmed | De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title_short | De novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
title_sort | de novo mapping of α-helix recognition sites on protein surfaces using unbiased libraries |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9907135/ https://www.ncbi.nlm.nih.gov/pubmed/36534810 http://dx.doi.org/10.1073/pnas.2210435119 |
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