Firefly luciferase mutants as sensors of proteome stress
Open accessAbstract
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.
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The aggregation quantification method was validated in animals expressing muscle-specific FlucDM-GFP.
“To analyze this process systematically, we developed a sensitive method for the quantification of aggregated proteins (see Experimental Procedures) and validated it in animals expressing muscle specific FlucDM-GFP, a conformationally unstable mutant of firefly luciferase fused to GFP (Gupta et al., 2011) (Figure S4A and B).”
From Widespread Proteome Remodeling and Aggregation in Aging C. elegans (2015) · cites this paper as Gupta et al., 2011
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The proteostasis network and its decline in ageing
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- Authors
- Gupta, R., Kasturi, P., Bracher, A., Loew, C., Zheng, M., Villella, A., Garza, D., Hartl, F. U., & Raychaudhuri, S.
- Affiliations
- Max Planck Institute of Biochemistry (DE)
- Proteostasis Therapeutics (United States) (US)
- Center for Integrated Protein Science Munich (DE)
- Corresponding author
- Hartl, F Ulrich
- Raychaudhuri, Swasti
- Venue
- Nature Methods
- Published
- 2011-09-04
- Volume
- 8
- Issue
- 10
- Pages
- 879-884
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2100680605
- Cited by
- 219
- References
- 439
- Citations by year
- 2026: 7 · 2025: 13 · 2024: 27 · 2023: 24 · 2022: 16 · 2021: 13 · 2020: 18 · 2019: 11 · 2018: 18 · 2017: 14 · 2016: 10 · 2015: 16 · 2014: 15 · 2013: 13 · 2012: 4
- FWCI
- 4.16
- Type
- journal article
- Language
- en
- Open access
- green
- hdl.handle.net
- Licence
- other-oa
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion, other-oa)
- Max Planck Institute for Plasma Physics (submittedVersion)
- Elektronische Hochschulschriften der LMU München (Ludwig-Maximilians-Universität München) (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
- Keywords
- firefly luciferase, proteostasis, protein aggregation, protein degradation, molecular chaperones, proteolytic enzymes, neurodegenerative diseases, aging, Caenorhabditis elegans, sensor proteins, protein solubility, protein misfolding
- MeSH terms
- HeLa Cells, Homeostasis, Humans, Models, Molecular, Proteins, Solubility, Proteome, Luciferases, Firefly, Luminescence, Green Fluorescent Proteins, Mutant Proteins
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