A Rapid Cellular FRET Assay of Polyglutamine Aggregation Identifies a Novel Inhibitor
| Autor: | Aye Aye K. Ma, J. Lawrence Marsh, Urvee A. Desai, Jieya Shao, Marc I. Diamond, Leslie M. Thompson, Judit Pallos, Sonia K Pollitt |
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| Jazyk: | angličtina |
| Předmět: |
Protein Folding
Pyridines Neuroscience(all) Drug Evaluation Preclinical Down-Regulation Nerve Tissue Proteins Biology Protein aggregation Protein Serine-Threonine Kinases Animals Genetically Modified 03 medical and health sciences 0302 clinical medicine Huntingtin Protein medicine Fluorescence Resonance Energy Transfer Animals Humans Nuclear protein Enzyme Inhibitors 030304 developmental biology Inclusion Bodies 0303 health sciences rho-Associated Kinases Cell Death General Neuroscience Neurodegeneration Intracellular Signaling Peptides and Proteins Nuclear Proteins Neurodegenerative Diseases medicine.disease Amides 3. Good health Cell biology Disease Models Animal Förster resonance energy transfer Drosophila melanogaster Biochemistry COS Cells Protein folding Biological Assay Photoreceptor Cells Invertebrate Signal transduction Peptides Trinucleotide Repeat Expansion 030217 neurology & neurosurgery Intracellular Signal Transduction |
| Zdroj: | Pollitt, SK; Pallos, J; Shao, J; Desai, UA; Ma, AAK; Thompson, LM; et al.(2003). A rapid cellular FRET assay of polyglutamine aggregation identifies a novel inhibitor. Neuron, 40(4), 685-694. doi: 10.1016/S0896-6273(03)00697-4. UC Irvine: Retrieved from: http://www.escholarship.org/uc/item/5457t1s5 |
| ISSN: | 0896-6273 |
| DOI: | 10.1016/S0896-6273(03)00697-4 |
| Popis: | Many neurodegenerative diseases, including tauopathies, Parkinson's disease, amyotrophic lateral sclerosis, and the polyglutamine diseases, are characterized by intracellular aggregation of pathogenic proteins. It is difficult to study modifiers of this process in intact cells in a high-throughput and quantitative manner, although this could facilitate molecular insights into disease pathogenesis. Here we introduce a high-throughput assay to measure intracellular polyglutamine protein aggregation using fluorescence resonance energy transfer (FRET). We screened over 2800 biologically active small molecules for inhibitory activity and have characterized one lead compound in detail. Y-27632, an inhibitor of the Rho-associated kinase p160ROCK, diminished polyglutamine protein aggregation (EC(50) congruent with 5 microM) and reduced neurodegeneration in a Drosophila model of polyglutamine disease. This establishes a novel high-throughput approach to study protein misfolding and aggregation associated with neurodegenerative diseases and implicates a signaling pathway of previously unrecognized importance in polyglutamine protein processing. |
| Databáze: | OpenAIRE |
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