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Methods Mol Biol. 2012;832:463-72. doi: 10.1007/978-1-61779-474-2_33.

Live-cell imaging of ubiquitin-proteasome system function.

Author information

1
Department of Biology, Stanford University, Stanford, CA, USA.

Abstract

The role of the ubiquitin-proteasome system (UPS) in maintaining protein homeostasis has generated a demand for assays that quantify UPS function in the presence of chemical and protein UPS inhibitors. Here, we describe protocols that measure changes in UPS reporter levels in response to changes in the expression level, localization, or aggregation state of a second protein. We utilize cell lines stably expressing fluorescent UPS substrates that are transfected with a second protein tagged with a compatible fluorophore. We describe protocols to correlate levels of UPS substrates with changes in the levels or properties of the transfected protein.

PMID:
22350906
PMCID:
PMC3711503
DOI:
10.1007/978-1-61779-474-2_33
[Indexed for MEDLINE]
Free PMC Article

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