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    Quantitative Analysis of α-Synuclein Solubility in Living Cells Using Split GFP Complementation

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    Author
    Kothawala, Ahmed; Kilpatrick, Kiri; Novoa, Jose Andres; Segatori, Laura
    Date
    2012
    Abstract
    Presently incurable, Parkinson's disease (PD) is the most common neurodegenerative movement disorder and affects 1% of the population over 60 years of age. The hallmarks of PD pathogenesis are the loss of dopaminergic neurons in the substantia nigra pars compacta, and the occurrence of proteinaceous cytoplasmic inclusions (Lewy bodies) in surviving neurons. Lewy bodies are mainly composed of the pre-synaptic protein alpha-synuclein (αsyn), an intrinsically unstructured, misfolding-prone protein with high propensity to aggregate. Quantifying the pool of soluble αsyn and monitoring αsyn aggregation in living cells is fundamental to study the molecular mechanisms of αsyn-induced cytotoxicity and develop therapeutic strategies to prevent αsyn aggregation. In this study, we report the use of a split GFP complementation assay to quantify αsyn solubility. Particularly, we investigated a series of naturally occurring and rationally designed αsyn variants and showed that this method can be used to study how αsyn sequence specificity affects its solubility. Furthermore, we demonstrated the utility of this assay to explore the influence of the cellular folding network on αsyn solubility. The results presented underscore the utility of the split GFP assay to quantify αsyn solubility in living cells.
    Citation
    Kothawala, Ahmed, Kilpatrick, Kiri, Novoa, Jose Andres, et al.. "Quantitative Analysis of α-Synuclein Solubility in Living Cells Using Split GFP Complementation." PLoS ONE, 7, no. 8 (2012) Public Library of Science: e43505. http://dx.doi.org/10.1371/journal.pone.0043505.
    Published Version
    http://dx.doi.org/10.1371/journal.pone.0043505
    Type
    Journal article
    Publisher
    Public Library of Science
    Citable link to this page
    https://hdl.handle.net/1911/78897
    Rights
    This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
    Link to License
    http://creativecommons.org/licenses/by/4.0/
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    Home | FAQ | Contact Us | Privacy Notice | Accessibility Statement
    Managed by the Digital Scholarship Services at Fondren Library, Rice University
    Physical Address: 6100 Main Street, Houston, Texas 77005
    Mailing Address: MS-44, P.O.BOX 1892, Houston, Texas 77251-1892
    Site Map