Site-Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover.
bio-orthogonal reactions
fluorescent probes
genetic code expansion
interleukins
protein folding
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
01 07 2020
01 07 2020
Historique:
received:
27
10
2019
revised:
28
01
2020
pubmed:
6
2
2020
medline:
8
6
2021
entrez:
4
2
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32011787
doi: 10.1002/cbic.201900651
pmc: PMC7383901
doi:
Substances chimiques
Amino Acids
0
Fluorescent Dyes
0
Interleukins
0
Protein Subunits
0
Amino Acyl-tRNA Synthetases
EC 6.1.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1861-1867Informations de copyright
© 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.
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