A Novel Destabilizing Domain Based on a Small-Molecule Dependent Fluorophore
Open accessAbstract
Tools that can directly regulate the activity of any protein-of-interest are valuable in the study of complex biological processes. Herein, we describe the development of a novel protein domain that exhibits small molecule-dependent stability and fluorescence based on the bilirubin-inducible fluorescent protein, UnaG. When genetically fused to any protein-of-interest, this fluorescent destabilizing domain (FDD) confers its instability to the entire fusion protein, facilitating the rapid degradation of the fusion. In the presence of its cognate ligand bilirubin (BR), the FDD fusion becomes stable and fluorescent. This new chemical genetic tool allows for rapid, reversible, and tunable control over the stability and fluorescence of a wide range of protein targets.
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The UnaG degron mutant (A36V, R136G) acts as a destabilizing degron on fusion proteins in the absence of bilirubin.
“Random mutagenesis and screening of UnaG yielded a two amino acid substitution mutant (A36V, R136G), which acts as a destabilizing degron on fusion proteins in the absence of BR (tested on mCherry, proteasome interactors zfand2a and zfand2b, ubiquitin conjugating enzyme ube2n and the cell cycle regulator p21) (Fig. 2b) [22].”
From Advances in targeted degradation of endogenous proteins (2019) · cites this paper as [22]
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- Authors
- Navarro, R., Chen, L.-C., Rakhit, R., & Wandless, T. J.
- Corresponding author
- Wandless, Thomas J.
- Venue
- ACS Chemical Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-06-06
- Volume
- 11
- Issue
- 8
- Pages
- 2101-2104
- ISSN
- 1554-8929 (ISSN-L)
- 1554-8937
- PMC
- PMC4992426
- OpenAlex
- W2409640222
- Cited by
- 31
- References
- 20
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 2 · 2023: 2 · 2022: 1 · 2021: 5 · 2020: 3 · 2019: 1 · 2018: 4 · 2017: 5 · 2016: 3
- FWCI
- 2.73
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- ACS Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Canadian Institutes of Health Research
- Division of Biological Infrastructure (1306670)
- National Institute of General Medical Sciences (GM073046)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Keywords
- UnaG, protein stability, chemical genetics, small-molecule ligands, protein degradation, protein fusion
- MeSH terms
- Animals, Fluorescent Dyes, Microscopy, Fluorescence, NIH 3T3 Cells, Mice
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