title: Quenched Hydrogen Exchange NMR of Amyloid Fibrils.
abstract: Amyloid fibrils are associated with a number of human diseases. These aggregatively misfolded intermolecular β-sheet assemblies constitute some of the most challenging targets in structural biology because to their complexity, size, and insolubility. Here, protocols and controls are described for experiments designed to study hydrogen-bonding in amyloid fibrils indirectly, by transferring information about amide proton occupancy in the fibrils to the dimethyl sulfoxide-denatured state. Since the denatured state is amenable to solution NMR spectroscopy, the method can provide residue-level-resolution data on hydrogen exchange for the monomers that make up the fibrils.
authors.affiliation: Department of Molecular and Cell Biology, University of Connecticut, Unit 3125, BSP 209, 91 N. Eagleville Rd., Storrs, CT, 06269-3125, USA. andrei.alexandrescu@uconn.edu.
authors.forename: Andrei T
authors.initials: AT
authors.lastname: Alexandrescu
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism; D003903:Deuterium / Q000737:chemistry; D006801:Humans; D006859:Hydrogen / Q000737:chemistry*; D006860:Hydrogen Bonding; D008279:Magnetic Resonance Imaging / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D017510:Protein Folding
chemical_list: D000682:Amyloid; D006859:Hydrogen; D003903:Deuterium
keywords: Aggregation; Amyloidogenic proteins; Hydrogen exchange; Hydrogen-bonding; NMR; Protein dynamics; Protein folding; Protein stability; Protein structure; Secondary structure