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Development of a DUB-selective fluorogenic substrate

Ubiquitination is a post-translational modification that is involved in a plethora of cellular processes. Target proteins can be specifically modified with a single ubiquitin (Ub) molecule or with complex chains. In recent years, research has focused on deubiquitinating enzymes (DUBs) as potential t...

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Autores principales: Gjonaj, Lorina, Sapmaz, Aysegul, Flierman, Dennis, Janssen, George M. C., van Veelen, Peter A., Ovaa, Huib
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988746/
https://www.ncbi.nlm.nih.gov/pubmed/32110315
http://dx.doi.org/10.1039/c9sc02226k
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author Gjonaj, Lorina
Sapmaz, Aysegul
Flierman, Dennis
Janssen, George M. C.
van Veelen, Peter A.
Ovaa, Huib
author_facet Gjonaj, Lorina
Sapmaz, Aysegul
Flierman, Dennis
Janssen, George M. C.
van Veelen, Peter A.
Ovaa, Huib
author_sort Gjonaj, Lorina
collection PubMed
description Ubiquitination is a post-translational modification that is involved in a plethora of cellular processes. Target proteins can be specifically modified with a single ubiquitin (Ub) molecule or with complex chains. In recent years, research has focused on deubiquitinating enzymes (DUBs) as potential therapeutic candidates in various diseases. USP16 is an emerging target due to its involvement in mitosis and stem cell self-renewal. Generally, activity-based probes (ABPs) used to study DUBs are based on the ubiquitin scaffold, thus lacking target selectivity. To overcome this issue, we designed a Ub-based activity probe bearing specific mutations to achieve selectivity for USP16, by combining structural modelling and analysis and mutational calculation predictions. We develop a fluorogenic substrate, the first of its kind, that is processed exclusively by USP16, which allows us to monitor USP16 activity in complex samples.
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spelling pubmed-69887462020-02-27 Development of a DUB-selective fluorogenic substrate Gjonaj, Lorina Sapmaz, Aysegul Flierman, Dennis Janssen, George M. C. van Veelen, Peter A. Ovaa, Huib Chem Sci Chemistry Ubiquitination is a post-translational modification that is involved in a plethora of cellular processes. Target proteins can be specifically modified with a single ubiquitin (Ub) molecule or with complex chains. In recent years, research has focused on deubiquitinating enzymes (DUBs) as potential therapeutic candidates in various diseases. USP16 is an emerging target due to its involvement in mitosis and stem cell self-renewal. Generally, activity-based probes (ABPs) used to study DUBs are based on the ubiquitin scaffold, thus lacking target selectivity. To overcome this issue, we designed a Ub-based activity probe bearing specific mutations to achieve selectivity for USP16, by combining structural modelling and analysis and mutational calculation predictions. We develop a fluorogenic substrate, the first of its kind, that is processed exclusively by USP16, which allows us to monitor USP16 activity in complex samples. Royal Society of Chemistry 2019-09-30 /pmc/articles/PMC6988746/ /pubmed/32110315 http://dx.doi.org/10.1039/c9sc02226k Text en This journal is © The Royal Society of Chemistry 2019 http://creativecommons.org/licenses/by-nc/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported Licence (CC BY-NC 3.0)
spellingShingle Chemistry
Gjonaj, Lorina
Sapmaz, Aysegul
Flierman, Dennis
Janssen, George M. C.
van Veelen, Peter A.
Ovaa, Huib
Development of a DUB-selective fluorogenic substrate
title Development of a DUB-selective fluorogenic substrate
title_full Development of a DUB-selective fluorogenic substrate
title_fullStr Development of a DUB-selective fluorogenic substrate
title_full_unstemmed Development of a DUB-selective fluorogenic substrate
title_short Development of a DUB-selective fluorogenic substrate
title_sort development of a dub-selective fluorogenic substrate
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988746/
https://www.ncbi.nlm.nih.gov/pubmed/32110315
http://dx.doi.org/10.1039/c9sc02226k
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