Back to Search
Start Over
Discovery of novel proteasome inhibitors using a high-content cell-based screening system
- Source :
- PLoS ONE, Vol 4, Iss 12, p e8503 (2009), PLoS ONE
- Publication Year :
- 2009
- Publisher :
- Public Library of Science (PLoS), 2009.
-
Abstract
- The regulated degradation of damaged or misfolded proteins, as well as down-regulation of key signaling proteins, within eukaryotic and bacterial cells is catalyzed primarily by large, ATP-dependent multimeric proteolytic complexes, termed proteasomes. Inhibition of proteasomal activity affects a wide variety of physiological and pathological processes, and was found to be particularly effective for cancer therapy. We report here on the development of a novel high throughput assay for proteasome inhibition using a unique, highly sensitive live-cell screening, based on the cytoplasm-to-nucleus translocation of a fluorescent proteasome inhibition reporter (PIR) protein, consisting of nuclear localization signal-deficient p53 derivative. We further show here that mdm2, a key negative regulator of p53 plays a key role in the accumulation of PIR in the nucleus upon proteasome inhibition. Using this assay, we have screened the NCI Diversity Set library, containing 1,992 low molecular weight synthetic compounds, and identified four proteasome inhibitors. The special features of the current screen, compared to those of other approaches are discussed.
- Subjects :
- Cell Survival
lcsh:Medicine
Biology
Protein Engineering
Biochemistry
Small Molecule Libraries
Proto-Oncogene Proteins c-mdm2
Genes, Reporter
Cell Line, Tumor
Drug Discovery
medicine
Humans
Enzyme Inhibitors
lcsh:Science
Cell Nucleus
Multidisciplinary
Drug discovery
lcsh:R
Biochemistry/Chemical Biology of the Cell
Reproducibility of Results
Cell Biology
Protein engineering
National Cancer Institute (U.S.)
United States
High-Throughput Screening Assays
Transport protein
Cell biology
Protein Transport
Cell nucleus
medicine.anatomical_structure
Proteasome
biology.protein
Mdm2
lcsh:Q
Biochemistry/Drug Discovery
Proteasome Inhibitors
Nuclear localization sequence
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 4
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....482c252b59d350173699162c1f7c465b