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Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover

Proteins that terminally fail to acquire their native structure are detected and degraded by cellular quality control systems. Insights into cellular protein quality control are key to a better understanding of how cells establish and maintain the integrity of their proteome and of how failures in t...

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Autores principales: Mideksa, Yonatan G., Fottner, Maximilian, Braus, Sebastian, Weiß, Caroline A. M., Nguyen, Tuan‐Anh, Meier, Susanne, Lang, Kathrin, Feige, Matthias J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7383901/
https://www.ncbi.nlm.nih.gov/pubmed/32011787
http://dx.doi.org/10.1002/cbic.201900651
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author Mideksa, Yonatan G.
Fottner, Maximilian
Braus, Sebastian
Weiß, Caroline A. M.
Nguyen, Tuan‐Anh
Meier, Susanne
Lang, Kathrin
Feige, Matthias J.
author_facet Mideksa, Yonatan G.
Fottner, Maximilian
Braus, Sebastian
Weiß, Caroline A. M.
Nguyen, Tuan‐Anh
Meier, Susanne
Lang, Kathrin
Feige, Matthias J.
author_sort Mideksa, Yonatan G.
collection PubMed
description Proteins that terminally fail to acquire their native structure are detected and degraded by cellular quality control systems. Insights into cellular protein quality control are key to a better understanding of how cells establish and maintain the integrity of their proteome and of how failures in these processes cause human disease. Here we have used genetic code expansion and fast bio‐orthogonal reactions to monitor protein turnover in mammalian cells through a fluorescence‐based assay. We have used immune signaling molecules (interleukins) as model substrates and shown that our approach preserves normal cellular quality control, assembly processes, and protein functionality and works for different proteins and fluorophores. We have further extended our approach to a pulse‐chase type of assay that can provide kinetic insights into cellular protein behavior. Taken together, this study establishes a minimally invasive method to investigate protein turnover in cells as a key determinant of cellular homeostasis.
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spelling pubmed-73839012020-07-27 Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover Mideksa, Yonatan G. Fottner, Maximilian Braus, Sebastian Weiß, Caroline A. M. Nguyen, Tuan‐Anh Meier, Susanne Lang, Kathrin Feige, Matthias J. Chembiochem Full Papers Proteins that terminally fail to acquire their native structure are detected and degraded by cellular quality control systems. Insights into cellular protein quality control are key to a better understanding of how cells establish and maintain the integrity of their proteome and of how failures in these processes cause human disease. Here we have used genetic code expansion and fast bio‐orthogonal reactions to monitor protein turnover in mammalian cells through a fluorescence‐based assay. We have used immune signaling molecules (interleukins) as model substrates and shown that our approach preserves normal cellular quality control, assembly processes, and protein functionality and works for different proteins and fluorophores. We have further extended our approach to a pulse‐chase type of assay that can provide kinetic insights into cellular protein behavior. Taken together, this study establishes a minimally invasive method to investigate protein turnover in cells as a key determinant of cellular homeostasis. John Wiley and Sons Inc. 2020-03-09 2020-07-01 /pmc/articles/PMC7383901/ /pubmed/32011787 http://dx.doi.org/10.1002/cbic.201900651 Text en © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Full Papers
Mideksa, Yonatan G.
Fottner, Maximilian
Braus, Sebastian
Weiß, Caroline A. M.
Nguyen, Tuan‐Anh
Meier, Susanne
Lang, Kathrin
Feige, Matthias J.
Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title_full Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title_fullStr Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title_full_unstemmed Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title_short Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
title_sort site‐specific protein labeling with fluorophores as a tool to monitor protein turnover
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7383901/
https://www.ncbi.nlm.nih.gov/pubmed/32011787
http://dx.doi.org/10.1002/cbic.201900651
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