Fast flow microfluidics and single-molecule fluorescence for the rapid characterization of α-synuclein oligomers.
Anal Chem
; 87(17): 8818-26, 2015 Sep 01.
Article
en En
| MEDLINE
| ID: mdl-26258431
ABSTRACT
α-Synuclein oligomers can be toxic to cells and may be responsible for cell death in Parkinson's disease. Their typically low abundance and highly heterogeneous nature, however, make such species challenging to study using traditional biochemical techniques. By combining fast-flow microfluidics with single-molecule fluorescence, we are able to rapidly follow the process by which oligomers of αS are formed and to characterize the species themselves. We have used the technique to show that populations of oligomers with different FRET efficiencies have varying stabilities when diluted into low ionic strength solutions. Interestingly, we have found that oligomers formed early in the aggregation pathway have electrostatic repulsions that are shielded in the high ionic strength buffer and therefore dissociate when diluted into lower ionic strength solutions. This property can be used to isolate different structural groups of αS oligomers and can help to rationalize some aspects of αS amyloid fibril formation.
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Bases de datos:
MEDLINE
Asunto principal:
Transferencia Resonante de Energía de Fluorescencia
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Técnicas Analíticas Microfluídicas
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Alfa-Sinucleína
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Fluorescencia
Idioma:
En
Revista:
Anal Chem
Año:
2015
Tipo del documento:
Article
País de afiliación:
Reino Unido