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A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes
The post-translational modification of proteins with ubiquitin (Ub) and Ub-like modifiers (Ubls) represents one of the most important regulators in eukaryotic biology. Polymeric Ub/Ubl chains of distinct topologies control the activity, stability, interaction and localization of almost all cellular...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8585875/ https://www.ncbi.nlm.nih.gov/pubmed/34764289 http://dx.doi.org/10.1038/s41467-021-26812-9 |
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author | Fottner, Maximilian Weyh, Maria Gaussmann, Stefan Schwarz, Dominic Sattler, Michael Lang, Kathrin |
author_facet | Fottner, Maximilian Weyh, Maria Gaussmann, Stefan Schwarz, Dominic Sattler, Michael Lang, Kathrin |
author_sort | Fottner, Maximilian |
collection | PubMed |
description | The post-translational modification of proteins with ubiquitin (Ub) and Ub-like modifiers (Ubls) represents one of the most important regulators in eukaryotic biology. Polymeric Ub/Ubl chains of distinct topologies control the activity, stability, interaction and localization of almost all cellular proteins and elicit a variety of biological outputs. Our ability to characterize the roles of distinct Ub/Ubl topologies and to identify enzymes and receptors that create, recognize and remove these modifications is however hampered by the difficulty to prepare them. Here we introduce a modular toolbox (Ubl-tools) that allows the stepwise assembly of Ub/Ubl chains in a flexible and user-defined manner facilitated by orthogonal sortase enzymes. We demonstrate the universality and applicability of Ubl-tools by generating distinctly linked Ub/Ubl hybrid chains, and investigate their role in DNA damage repair. Importantly, Ubl-tools guarantees straightforward access to target proteins, site-specifically modified with distinct homo- and heterotypic (including branched) Ub chains, providing a powerful approach for studying the functional impact of these complex modifications on cellular processes. |
format | Online Article Text |
id | pubmed-8585875 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-85858752021-11-15 A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes Fottner, Maximilian Weyh, Maria Gaussmann, Stefan Schwarz, Dominic Sattler, Michael Lang, Kathrin Nat Commun Article The post-translational modification of proteins with ubiquitin (Ub) and Ub-like modifiers (Ubls) represents one of the most important regulators in eukaryotic biology. Polymeric Ub/Ubl chains of distinct topologies control the activity, stability, interaction and localization of almost all cellular proteins and elicit a variety of biological outputs. Our ability to characterize the roles of distinct Ub/Ubl topologies and to identify enzymes and receptors that create, recognize and remove these modifications is however hampered by the difficulty to prepare them. Here we introduce a modular toolbox (Ubl-tools) that allows the stepwise assembly of Ub/Ubl chains in a flexible and user-defined manner facilitated by orthogonal sortase enzymes. We demonstrate the universality and applicability of Ubl-tools by generating distinctly linked Ub/Ubl hybrid chains, and investigate their role in DNA damage repair. Importantly, Ubl-tools guarantees straightforward access to target proteins, site-specifically modified with distinct homo- and heterotypic (including branched) Ub chains, providing a powerful approach for studying the functional impact of these complex modifications on cellular processes. Nature Publishing Group UK 2021-11-11 /pmc/articles/PMC8585875/ /pubmed/34764289 http://dx.doi.org/10.1038/s41467-021-26812-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Fottner, Maximilian Weyh, Maria Gaussmann, Stefan Schwarz, Dominic Sattler, Michael Lang, Kathrin A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title | A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title_full | A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title_fullStr | A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title_full_unstemmed | A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title_short | A modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
title_sort | modular toolbox to generate complex polymeric ubiquitin architectures using orthogonal sortase enzymes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8585875/ https://www.ncbi.nlm.nih.gov/pubmed/34764289 http://dx.doi.org/10.1038/s41467-021-26812-9 |
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