title: Morphology and antibody recognition of synthetic beta-amyloid peptides.
abstract: To elucidate the relationship between amyloid fibril formation in Alzheimer disease (AD) and the primary structure of the beta-amyloid protein (beta-AP), we investigated the ability of peptides sharing sequences with beta-AP to form fibrils in vitro and to recognize anti-beta-amyloid antisera. The peptides, which were synthesized using a FMOC solid phase procedure and purified by HPLC, consisted of residues 6-25 from the putative aqueous domain, residues 22-35, which overlaps the putative aqueous and transmembrane domains, and residues 1-38 and 1-40 representing nearly the full length of beta-AP. Electron microscopy of negative-stained or thin-sectioned preparations revealed that the peptides assembled into fibrils having different morphologies, some of which resembled in situ AD amyloid. Peptide 6-25 fibrils had diameters of 50-80 A and occasionally showed a central groove suggestive of constituent filaments. Cross sections of the fibril showed a penta- or hexameric arrangement of globular subunits with diameters of 25-30 A. Peptide 22-35 fibrils were helical, with a pitch of 1,100 A and a width of 120 A at its greatest and 50-60 A at its narrowest. The fibrils formed by peptides 1-38 and 1-40 were 70-90 A in diameter. When the peptide assemblies were singly oriented by sedimentation or doubly oriented in a magnetic field, their X-ray diffraction patterns all showed reflections typical of a cross-beta pleated sheet conformation. The patterns differed mainly in their small-angle equatorial intensity, which arises from the packing of fibrils having different widths. Antiserum raised to either native amyloid or to synthetic peptide beta-(1-28) was highly reactive in an inhibition-ELISA assay to beta-(6-25) and beta-(1-38), but not to beta-(22-35), and immunostained beta-(1-40) on Western blots. These studies show that the beta-(6-25), beta-(1-38) and beta-(1-40) peptides can assemble into cross-beta fibrils that retain epitopes characteristic of AD amyloid.
authors.affiliation: Children's Hospital, Department of Neurology, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: P E
authors.initials: PE
authors.lastname: Fraser
authors.forename: L K
authors.initials: LK
authors.lastname: Duffy
authors.forename: M B
authors.initials: MB
authors.lastname: O'Malley
authors.forename: J
authors.initials: J
authors.lastname: Nguyen
authors.forename: H
authors.initials: H
authors.lastname: Inouye
authors.forename: D A
authors.initials: DA
authors.lastname: Kirschner
mesh_terms: D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000032:analysis* / Q000276:immunology; D016564:Amyloid beta-Protein Precursor; D000937:Antigen-Antibody Reactions; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007120:Immunochemistry; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011498:Protein Precursors / Q000032:analysis* / Q000276:immunology; D014961:X-Ray Diffraction
chemical_list: D016229:Amyloid beta-Peptides; D016564:Amyloid beta-Protein Precursor; D011498:Protein Precursors