Impact of linker length on the activity of PROTACs
Open accessAbstract
Conventional genetic approaches have provided a powerful tool in the study of proteins. However, these techniques often preclude selective manipulation of temporal and spatial protein functions, which is crucial for the investigation of dynamic cellular processes. To overcome these limitations, a small molecule-based novel technology termed “PROteolysis TArgeting ChimeraS (PROTACs)” has been developed, targeting proteins for degradation at the post-translational level. Despite the promising potential of PROTACs to serve as molecular probes of complex signaling pathways, their design has not been generalized for broad application. Here, we present the first generalized approach for PROTAC design by fine-tuning the distance between the two participating partner proteins, the E3 ubiquitin ligase and the target protein. As such, we took a chemical approach to create estrogen receptor(ER)-α targeting PROTACs with varying linker lengths and the loss of the ER in cultured cells was monitored via western blot and fluorometric analyses. We found a significant effect of chain length on PROTAC efficacy, and, in this case, the optimum distance between the E3 recognition motif and the ligand was a 16 atom chain length. The information gathered from this experiment may offer a generalizable PROTAC design strategy to further the expansion of the PROTAC toolbox, opening new possibilities for the broad application of the PROTAC strategy in the study of multiple signaling pathways.
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The linker can contribute to PROTAC solution conformation and degradation efficiency (DC50/Dmax), and influence E3 ligase-POI interactions and presentation of the POI within the zone of ubiquitylation.
“The linker can contribute to the PROTAC solution conformation68 and degradation efficiency (DC50/Dmax)69, and influence the E3 ligase–POI interactions and presentation of the POI within the zone of ubiquitylation (Fig. 5).”
From PROTAC targeted protein degraders: the past is prologue (2022) · cites this paper as [69]
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- Authors
- Cyrus, K., Wehenkel, M., Choi, E.-Y., Han, H.-J., Lee, H., Swanson, H., & Kim, K.-B.
- Affiliations
- University of Kentucky (US)
- Department of Pharmaceutical Sciences, University of Kentucky a , Lexington, Kentucky, 40536-0596, USA kbkim2@uky.edu +1 859 257 7564 +1 859 257 5301
- Venue
- Molecular BioSystems
- Publisher
- Oxford University Press (OUP)
- Published
- 2010-10-04
- Volume
- 7
- Issue
- 2
- Pages
- 359-364
- ISSN
- 1742-2051 (ISSN-L)
- 1742-206X
- PMC
- PMC3835402
- OpenAlex
- W2002111767
- Cited by
- 232
- References
- 26
- Citations by year
- 2026: 27 · 2025: 39 · 2024: 36 · 2023: 23 · 2022: 36 · 2021: 24 · 2020: 18 · 2019: 9 · 2018: 4 · 2017: 6 · 2016: 4 · 2015: 1 · 2013: 1 · 2012: 2
- FWCI
- 0.65
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular BioSystems (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- American Foundation for Pharmaceutical Education
- National Institutes of Health
- National Cancer Institute (R01CA131059, CA131059)
- National Institute of Environmental Health Sciences (ES014849, R01ES014849)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- PROTACs, linker length, E3 ubiquitin ligase, estrogen receptor alpha, protein degradation, PROTAC design
- MeSH terms
- Humans, Hydrolysis, Proteins, Ubiquitin-Protein Ligases, Cell Line, Tumor, Estrogen Receptor alpha
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