title: Interactions between amyloidophilic dyes and their relevance to studies of amyloid inhibitors.
abstract: Amyloid fibrils are filamentous aggregates of peptides and proteins implicated in a range of neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. It has been known almost since their discovery that these β-sheet-rich proteinacious assemblies bind a range of specific dyes that, combined with other biophysical techniques, are convenient probes of the process of amyloid fibril formation. Two prominent examples of such dyes are Congo red (CR) and Thioflavin T (ThT). It has been reported that in addition to having a diagnostic role, CR is an inhibitor of the formation of amyloid structures, and these two properties have both been explained in terms of the same specific noncovalent interactions between the fibrils and the dye molecules. In this article, we show by means of quartz-crystal microbalance measurements that the binding of both ThT and CR to amyloid fibrils formed by the peptide whose aggregation is associated with Alzheimer's disease, Aβ(1-42), can be directly observed, and that the presence of CR interferes with the binding of ThT. Light scattering and fluorescence measurements confirm that an interaction exists between these dyes that can interfere with their ability to reflect accurately the quantity of amyloid material present in a given sample. Furthermore, we show that CR does not inhibit the process of amyloid fibril elongation, and therefore demonstrate the ability of the quartz-crystal microbalance method not only to detect and study the binding of small molecules to amyloid fibrils, but also to elucidate the mode of action of potential inhibitors.
authors.affiliation: Department of Engineering, University of Cambridge, Nanoscience Center, Cambridge, UK.
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Buell
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.forename: Tuomas P J
authors.initials: TP
authors.lastname: Knowles
authors.forename: Mark E
authors.initials: ME
authors.lastname: Welland
mesh_terms: D000682:Amyloid / Q000378:metabolism* / Q000648:ultrastructure; D052160:Benzothiazoles; D004396:Coloring Agents / Q000378:metabolism*; D003224:Congo Red / Q000378:metabolism*; D006801:Humans; D008027:Light; D010316:Particle Size; D058525:Quartz Crystal Microbalance Techniques; D012542:Scattering, Radiation; D013844:Thiazoles / Q000378:metabolism*
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D004396:Coloring Agents; D013844:Thiazoles; C009462:thioflavin T; D003224:Congo Red