期刊
EXPERT OPINION ON THERAPEUTIC TARGETS
卷 19, 期 5, 页码 589-603出版社
INFORMA HEALTHCARE
DOI: 10.1517/14728222.2015.1009448
关键词
alpha-synuclein; high-throughput screening; oligomers; Parkinson's disease; protein-fragment complementation; synucleinopathy
资金
- Mayo Clinic/SBMRI
- Florida Translational Research Program (FTRP)
- NIH [NS063963]
Objective: Reducing the burden of alpha-synuclein oligomeric species represents a promising approach for disease-modifying therapies against synucleinopathies such as Parkinson's disease and dementia with Lewy bodies. However, the lack of efficient drug discovery strategies that specifically target alpha-synuclein oligomers has been a limitation to drug discovery programs. Research design and methods: Here we describe an innovative strategy that harnesses the power of bimolecular protein-fragment complementation to monitor synuclein-synuclein interactions. We have developed two robust models to monitor alpha-synuclein oligomerization by generating novel stable cell lines expressing alpha-synuclein fusion proteins for either fluorescent or bioluminescent protein-fragment complementation under the tetracycline-controlled transcriptional activation system. Main outcome measures: A pilot screen was performed resulting in the identification of two potential hits, a p38 MAPK inhibitor and a casein kinase 2 inhibitor, thereby demonstrating the suitability of our protein-fragment complementation assay for the measurement of alpha-synuclein oligomerization in living cells at high throughput. Conclusions: The application of the strategy described herein to monitor alpha-synuclein oligomer formation in living cells with high throughput will facilitate drug discovery efforts for disease-modifying therapies against synucleinopathies and other proteinopathies.
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