title: Fluorescence Methods for Unraveling Oligomeric Amyloid Intermediates.
abstract: Amyloid oligomers are considered to be the relevant toxic species in many amyloid diseases and much research effort has been devoted to fully characterize these oligomers. Despite their importance, oligomers have proven to be difficult to characterize structurally. Information on their aggregation number is scarce, largely because standard techniques struggle to provide reliable results. In this chapter, we present two different methods that reproducibly yield fluorescently labeled α-Synuclein oligomers. We then discuss a new approach, combining single-molecule photobleaching and sub-stoichiometric fluorescent labeling, that we have developed to determine the aggregation number of supramolecular protein assemblies.
authors.affiliation: FOM Institute AMOLF, Science Park 104, 1098 XG, Amsterdam, The Netherlands.|Nanobiophysics, MESA+ Institute for Nanotechnology, Faculty of Science and Technology, University of Twente, 217, 7500 AE, Enschede, The Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Zijlstra
authors.affiliation: Nanobiophysics, MESA+ Institute for Nanotechnology, Faculty of Science and Technology, University of Twente, 217, 7500 AE, Enschede, The Netherlands.|Nanobiophysics, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 217, 7500 AE, Enschede, The Netherlands.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Schilderink
authors.affiliation: FOM Institute AMOLF, Science Park 104, 1098 XG, Amsterdam, The Netherlands. subramaniam@amolf.nl.|Nanobiophysics, MESA+ Institute for Nanotechnology, Faculty of Science and Technology, University of Twente, 217, 7500 AE, Enschede, The Netherlands. subramaniam@amolf.nl.|Nanobiophysics, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, 217, 7500 AE, Enschede, The Netherlands. subramaniam@amolf.nl.
authors.forename: Vinod
authors.initials: V
authors.lastname: Subramaniam
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D010300:Parkinson Disease / Q000235:genetics; D055503:Protein Multimerization / Q000235:genetics*; D013194:Staining and Labeling / Q000379:methods*; D051844:alpha-Synuclein / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D005456:Fluorescent Dyes; D051844:alpha-Synuclein
keywords: Oligomer; Photobleaching; Single molecule; Sub-stoichiometric; Supramolecular assembly; α-Synuclein