Elucidating the aggregation number of dopamine-induced α-synuclein oligomeric assemblies

Niels Zijlstra, Mireille M A E Claessens, Christian Blum, Vinod Subramaniam

    Research output: Contribution to JournalArticleAcademicpeer-review

    Abstract

    Conventional methods to determine the aggregation number, that is, the number of monomers per oligomer, struggle to yield reliable results for large protein aggregates, such as amyloid oligomers. We have previously demonstrated the use of a combination of single-molecule photobleaching and substoichiometric fluorescent labeling to determine the aggregation number of oligomers of human α-synuclein, implicated in Parkinson's disease. We show here that this approach is capable of accurately resolving mixtures of multiple distinct molecular species present in the same sample of dopamine-induced α-synuclein oligomers, and that we can determine the respective aggregation numbers of each species from a single histogram of bleaching steps. We found two distinct species with aggregation numbers of 15-19 monomers and 34-38 monomers. These results show that this single-molecule approach allows for the systematic study of the aggregation numbers of complex supramolecular assemblies formed under different aggregation conditions.

    Original languageEnglish
    Pages (from-to)440-6
    Number of pages7
    JournalBiophysical Journal
    Volume106
    Issue number2
    DOIs
    Publication statusPublished - 21 Jan 2014

    Keywords

    • Dopamine
    • Fluorescent Dyes
    • Humans
    • Photobleaching
    • Protein Multimerization
    • Protein Structure, Quaternary
    • alpha-Synuclein
    • Journal Article
    • Research Support, Non-U.S. Gov't

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