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Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes

The proteasome is a multi-catalytic molecular machine that plays a key role in the degradation of many cytoplasmic and nuclear proteins. The proteasome is essential and proteasome malfunction is associated with various disease pathologies. Proteasome activity depends on its catalytic subunits which...

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Autores principales: Schipper-Krom, Sabine, Sanz, Alicia Sanz, van Bodegraven, Emma J., Speijer, Dave, Florea, Bogdan I., Ovaa, Huib, Reits, Eric A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6710370/
https://www.ncbi.nlm.nih.gov/pubmed/31482094
http://dx.doi.org/10.3389/fmolb.2019.00056
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author Schipper-Krom, Sabine
Sanz, Alicia Sanz
van Bodegraven, Emma J.
Speijer, Dave
Florea, Bogdan I.
Ovaa, Huib
Reits, Eric A.
author_facet Schipper-Krom, Sabine
Sanz, Alicia Sanz
van Bodegraven, Emma J.
Speijer, Dave
Florea, Bogdan I.
Ovaa, Huib
Reits, Eric A.
author_sort Schipper-Krom, Sabine
collection PubMed
description The proteasome is a multi-catalytic molecular machine that plays a key role in the degradation of many cytoplasmic and nuclear proteins. The proteasome is essential and proteasome malfunction is associated with various disease pathologies. Proteasome activity depends on its catalytic subunits which are interchangeable and also on the interaction with the associated regulatory cap complexes. Here, we describe and compare various methods that allow the study of proteasome function in living cells. Methods include the use of fluorescently tagged proteasome subunits and the use of activity-based proteasome probes. These probes can be used in both biochemical assays and in microscopy-based experiments. Together with tagged proteasomes, they can be used to study proteasome localization, dynamics, and activity.
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spelling pubmed-67103702019-09-03 Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes Schipper-Krom, Sabine Sanz, Alicia Sanz van Bodegraven, Emma J. Speijer, Dave Florea, Bogdan I. Ovaa, Huib Reits, Eric A. Front Mol Biosci Molecular Biosciences The proteasome is a multi-catalytic molecular machine that plays a key role in the degradation of many cytoplasmic and nuclear proteins. The proteasome is essential and proteasome malfunction is associated with various disease pathologies. Proteasome activity depends on its catalytic subunits which are interchangeable and also on the interaction with the associated regulatory cap complexes. Here, we describe and compare various methods that allow the study of proteasome function in living cells. Methods include the use of fluorescently tagged proteasome subunits and the use of activity-based proteasome probes. These probes can be used in both biochemical assays and in microscopy-based experiments. Together with tagged proteasomes, they can be used to study proteasome localization, dynamics, and activity. Frontiers Media S.A. 2019-08-20 /pmc/articles/PMC6710370/ /pubmed/31482094 http://dx.doi.org/10.3389/fmolb.2019.00056 Text en Copyright © 2019 Schipper-Krom, Sanz, van Bodegraven, Speijer, Florea, Ovaa and Reits. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Schipper-Krom, Sabine
Sanz, Alicia Sanz
van Bodegraven, Emma J.
Speijer, Dave
Florea, Bogdan I.
Ovaa, Huib
Reits, Eric A.
Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title_full Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title_fullStr Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title_full_unstemmed Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title_short Visualizing Proteasome Activity and Intracellular Localization Using Fluorescent Proteins and Activity-Based Probes
title_sort visualizing proteasome activity and intracellular localization using fluorescent proteins and activity-based probes
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6710370/
https://www.ncbi.nlm.nih.gov/pubmed/31482094
http://dx.doi.org/10.3389/fmolb.2019.00056
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