title: Structure-selective anisotropy assay for amyloid Beta oligomers.
abstract: Amyloid β (Abeta) peptides in their oligomeric form have been proposed as major toxic species in Alzheimer's disease (AD). There are a limited number of anti-Abeta antibodies specific to oligomeric forms of Abeta compared to the monomeric form, and accurate measurement of oligomeric forms in biological samples, cerebrospinal fluid (CSF), or brain extracts remains challenging. We introduce an oligomer-specific (in preference to monomers or fibrils) fluorescence assay based on a conformationally sensitive bis-pyrene-labeled peptide that contains amino acid residues 16-35 of the human amyloid beta protein (pronucleon peptide, PP). This peptide exhibits a shift in fluorescence emission from pyrene excimer to pyrene monomer emission resulting from a conformational change. Specific binding of PP to oligomeric forms of Abeta can be monitored in solution by a change in fluorescence spectrum as well as a change in pyrene monomer fluorescence anisotropy (or polarization). The mechanism of binding and its relation to anisotropy and fluorescence lifetime changes are discussed. The development of a simple, rapid, anisotropy assay for measurement of Abeta oligomers is important for further study of the oligomers' role in AD, and specific detection of oligomers in biological samples, such as cerebrospinal fluid.
authors.affiliation: University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, Baltimore, Maryland 21201, United States; Adlyfe, Inc., Rockville, Maryland 20850, United States. evgenia_matveeva@hotmail.com
authors.forename: Evgenia G
authors.initials: EG
authors.lastname: Matveeva
authors.forename: Alan
authors.initials: A
authors.lastname: Rudolph
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Moll
authors.forename: Richard B
authors.initials: RB
authors.lastname: Thompson
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry*; D016880:Anisotropy; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry*; D010446:Peptide Fragments / Q000737:chemistry; D011721:Pyrenes; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D016229:Amyloid beta-Peptides; D009842:Oligopeptides; D010446:Peptide Fragments; D011721:Pyrenes