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Nat Methods. 2011 Sep 4;8(10):879-84. doi: 10.1038/nmeth.1697.

Firefly luciferase mutants as sensors of proteome stress.

Author information

1
Department of Cellular Biochemistry, Max Planck Institute of Biochemistry, Martinsried, Germany.

Abstract

Maintenance of cellular protein homeostasis (proteostasis) depends on a complex network of molecular chaperones, proteases and other regulatory factors. Proteostasis deficiency develops during normal aging and predisposes individuals for many diseases, including neurodegenerative disorders. Here we describe sensor proteins for the comparative measurement of proteostasis capacity in different cell types and model organisms. These sensors are increasingly structurally destabilized versions of firefly luciferase. Imbalances in proteostasis manifest as changes in sensor solubility and luminescence activity. We used EGFP-tagged constructs to monitor the aggregation state of the sensors and the ability of cells to solubilize or degrade the aggregated proteins. A set of three sensor proteins serves as a convenient toolkit to assess the proteostasis status in a wide range of experimental systems, including cell and organism models of stress, neurodegenerative disease and aging.

PMID:
21892152
DOI:
10.1038/nmeth.1697
[Indexed for MEDLINE]

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