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Development of a Cell-Based AlphaLISA Assay for High-Throughput Screening for Small Molecule Proteasome Modulators.
| Content Provider | Europe PMC |
|---|---|
| Author | Staerz, SophiaD. Lisabeth, Erika M. Njomen, Evert Dexheimer, Thomas S. Neubig, Richard R. Tepe, Jetze J. |
| Copyright Year | 2023 |
| Abstract | The balance between protein degradation and protein synthesisisa highly choreographed process generally called proteostasis. Mostintracellular protein degradation occurs through the ubiquitin–proteasomesystem (UPS). This degradation takes place through either a ubiquitin-dependentor a ubiquitin-independent proteasomal pathway. The ubiquitin-independentpathway selectively targets unfolded proteins, including intrinsicallydisordered proteins (IDPs). Dysregulation of proteolysis can leadto the accumulation of IDPs, seen in the pathogenesis of various diseases,including cancer and neurodegeneration. Therefore, the enhancementof the proteolytic activity of the 20S proteasome using small moleculeshas been identified as a promising pathway to combat IDP accumulation.Currently, there are a limited number of known small molecules thatenhance the activity of the 20S proteasome, and few are observed toexhibit enhanced proteasome activity in cell culture. Herein, we describethe development of a high-throughput screening assay to identify cell-permeableproteasome enhancers by utilizing an AlphaLISA platform that measuresthe degradation of a GFP conjugated intrinsically disordered protein,ornithine decarboxylase (ODC). Through the screening of the Prestwickand NIH Clinical Libraries, a kinase inhibitor, erlotinib, was identifiedas a new 20S proteasome enhancer, which enhances the degradation ofODC in cells and α-synuclein in vitro. |
| Journal | ACS Omega |
| Volume Number | 8 |
| PubMed Central reference number | PMC10157846 |
| Issue Number | 17 |
| PubMed reference number | 37151549 |
| e-ISSN | 24701343 |
| DOI | 10.1021/acsomega.3c01158 |
| Language | English |
| Publisher | American Chemical Society |
| Publisher Date | 2023-04-21 |
| Access Restriction | Open |
| Rights License | Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). © 2023 The Authors. Published by American Chemical Society |
| Content Type | Text |
| Resource Type | Article |
| Subject | Chemistry Chemical Engineering |
