Site‐Specific Protein Labeling with Fluorophores as a Tool To Monitor Protein Turnover
- Journal Article
Rights / licenseCreative Commons Attribution-NonCommercial 4.0 International
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. Show more
Journal / seriesChemBioChem
Pages / Article No.
Subjectbio-orthogonal reactions; fluorescent probes; genetic code expansion; interleukins; protein folding
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