title: Kinetic analysis of beta-amyloid fibril elongation.
abstract: We used surface plasmon resonance biosensors to evaluate the kinetics associated with the initial events of beta-amyloid (Abeta) fibril elongation. Fibrils were immobilized on the sensor chip surface and extended by exposure to soluble Abeta(1-40) peptide. The fibril surfaces bound Congo red, a marker for beta sheet structures, and exhibited a slow linear background decay that is consistent with fibril depolymerization. Sonicated fibrils supported elongation better than unsonicated fibrils, which is consistent with fibril extension reactions. The kinetic data revealed that peptide association and dissociation occurred in multiple steps. Kinetic rate constants for fibril extension were determined by globally fitting the response data with a three-step polymerization model. In the first step, the soluble peptide binds to the growing fibril tip in a readily reversible reaction. The subsequent steps likely allow bound peptide to be stabilized into the growing fiber through postbinding transitional events. Using a mutant peptide, F19P Abeta(1-40), we illustrate how the biosensor assay can be used to probe structure/function relationships of fibril elongation.
authors.affiliation: Center for Biomolecular Interaction Analysis, University of Utah, Salt Lake City, UT 84132, USA.
authors.forename: Michelle J
authors.initials: MJ
authors.lastname: Cannon
authors.forename: Angela D
authors.initials: AD
authors.lastname: Williams
authors.forename: Ronald
authors.initials: R
authors.lastname: Wetzel
authors.forename: David G
authors.initials: DG
authors.lastname: Myszka
mesh_terms: D016229:Amyloid beta-Peptides / Q000235:genetics / Q000378:metabolism*; D015374:Biosensing Techniques; D003224:Congo Red / Q000378:metabolism; D007700:Kinetics; D009154:Mutation; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D013010:Sonication; D020349:Surface Plasmon Resonance; D013997:Time Factors
chemical_list: D016229:Amyloid beta-Peptides; D010446:Peptide Fragments; C078592:amyloid beta-protein (1-40); D003224:Congo Red