title: Preparation of Amyloid Fibrils Seeded from Brain and Meninges.
abstract: Seeding of amyloid fibrils into fresh solutions of the same peptide or protein in disaggregated form leads to the formation of replicate fibrils, with close structural similarity or identity to the original fibrillar seeds. Here we describe procedures for isolating fibrils composed mainly of β-amyloid (Aβ) from human brain and from leptomeninges, a source of cerebral blood vessels, for investigating Alzheimer's disease and cerebral amyloid angiopathy. We also describe methods for seeding isotopically labeled, disaggregated Aβ peptide solutions for study using solid-state NMR and other techniques. These methods should be applicable to other types of amyloid fibrils, to Aβ fibrils from mice or other species, tissues other than brain, and to some non-fibrillar aggregates. These procedures allow for the examination of authentic amyloid fibrils and other protein aggregates from biological tissues without the need for labeling the tissue.
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, AMB N314, 5841 South Maryland Avenue, Chicago, IL, 60637, USA.
authors.forename: Kathryn P
authors.initials: KP
authors.lastname: Scherpelz
authors.affiliation: Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jun-Xia
authors.initials: JX
authors.lastname: Lu
authors.affiliation: Laboratory of Chemical Physics, NIDDK, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Tycko
authors.affiliation: Department of Biochemistry and Molecular Biology, The University of Chicago, AMB N314, 5841 South Maryland Avenue, Chicago, IL, 60637, USA. scmeredi@uchicago.edu.|Department of Pathology, The University of Chicago, Chicago, IL, 60637, USA. scmeredi@uchicago.edu.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Meredith
mesh_terms: D000544:Alzheimer Disease / Q000378:metabolism / Q000473:pathology; D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D016229:Amyloid beta-Peptides / Q000737:chemistry / Q000648:ultrastructure; D000818:Animals; D001921:Brain / Q000378:metabolism / Q000648:ultrastructure; D001923:Brain Chemistry; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008578:Meninges / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D018625:Microscopy, Atomic Force / Q000379:methods
chemical_list: D000682:Amyloid; D016229:Amyloid beta-Peptides
keywords: Alzheimer’s disease; Cerebral amyloid angiopathy; Fibril polymorphism; Fibril structure; Neuritic plaques; Protein aggregation; Protein aggregation diseases; Solid-state NMR; β-Amyloid