Elucidating the Aggregation Number of Dopamine-Induced alpha-Synuclein Oligomeric Assemblies

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

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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)440-446
Number of pages7
JournalBiophysical journal
Volume106
Issue number2
DOIs
Publication statusPublished - 2014

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Synucleins
alpha-Synuclein
Dopamine
Photobleaching
Amyloid
Parkinson Disease

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title = "Elucidating the Aggregation Number of Dopamine-Induced alpha-Synuclein Oligomeric Assemblies",
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",
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Elucidating the Aggregation Number of Dopamine-Induced alpha-Synuclein Oligomeric Assemblies. / Zijlstra, Niels; Claessens, Mireille M.A.E.; Blum, Christian; Subramaniam, Vinod.

In: Biophysical journal, Vol. 106, No. 2, 2014, p. 440-446.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Zijlstra, Niels

AU - Claessens, Mireille M.A.E.

AU - Blum, Christian

AU - Subramaniam, Vinod

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