A rapid cellular FRET assay of polyglutamine aggregation identifies a novel inhibitor

Sonia K. Pollitt, Judit Pallos, Jieya Shao, Urvee A. Desai, Aye Aye K Ma, Leslie Michels Thompson, J. Lawrence Marsh, Marc I. Diamond

Research output: Contribution to journalArticle

121 Scopus citations

Abstract

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 (EC50 ≅ 5 μM) 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.

Original languageEnglish (US)
Pages (from-to)685-694
Number of pages10
JournalNeuron
Volume40
Issue number4
DOIs
StatePublished - Nov 13 2003

ASJC Scopus subject areas

  • Neuroscience(all)

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    Pollitt, S. K., Pallos, J., Shao, J., Desai, U. A., Ma, A. A. K., Thompson, L. M., Marsh, J. L., & Diamond, M. I. (2003). A rapid cellular FRET assay of polyglutamine aggregation identifies a novel inhibitor. Neuron, 40(4), 685-694. https://doi.org/10.1016/S0896-6273(03)00697-4