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Development of a Cell-Based AlphaLISA Assay for High-Throughput Screening for Small Molecule Proteasome Modulators.
Sophia D Staerz1, Erika M Lisabeth1, Evert Njomen1
1Department of Chemistry, Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Michigan 48824, United States.
ACS Omega
|May 8, 2023
Summary
Researchers developed a new assay to find drugs that boost proteasome activity, crucial for clearing harmful proteins. Erlotinib, a kinase inhibitor, was identified as a promising proteasome enhancer, aiding in the degradation of disease-associated proteins.
Area of Science:
- Biochemistry
- Molecular Biology
- Cellular Biology
Background:
- Proteostasis, the balance between protein synthesis and degradation, is vital for cellular health.
- The ubiquitin-proteasome system (UPS) is the primary pathway for intracellular protein degradation.
- Dysregulation of proteolysis, particularly the accumulation of intrinsically disordered proteins (IDPs), is implicated in diseases like cancer and neurodegeneration.
Purpose of the Study:
- To develop a high-throughput screening assay for identifying cell-permeable 20S proteasome enhancers.
- To discover novel small molecules that enhance the degradation of intrinsically disordered proteins (IDPs).
Main Methods:
- Development of an AlphaLISA-based high-throughput screening assay.
- Assay utilized a GFP-conjugated intrinsically disordered protein, ornithine decarboxylase (ODC), to measure proteasome activity.
- Screening of the Prestwick and NIH Clinical Libraries.
Main Results:
- Identification of erlotinib, a known kinase inhibitor, as a novel 20S proteasome enhancer.
- Erlotinib demonstrated enhanced degradation of ODC in cell culture.
- Erlotinib also enhanced the degradation of alpha-synuclein *in vitro*.
Conclusions:
- The developed assay is effective for identifying proteasome enhancers.
- Erlotinib represents a promising therapeutic candidate for diseases associated with IDP accumulation.
- Targeting the 20S proteasome offers a potential strategy to combat proteostasis-related disorders.
