Structure of beta-crystallite assemblies formed by Alzheimer beta-amyloid protein analogues: analysis by x-ray diffraction

• Published in: Biophysical journal Vol. 64; no. 2; pp. 502 - 519
• Main Authors: Inouye, H., Fraser, P.E., Kirschner, D.A.
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Elsevier Inc 01.02.1993; Biophysical Society; The Biophysical Society
• Subjects: Amino Acid Sequence; Amyloid beta-Peptides - chemistry; Biological and medical sciences; Biophysical Phenomena; Biophysics; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; Electrons; Humans; Macromolecular Substances; Medical sciences; Models, Chemical; Models, Molecular; Molecular Sequence Data; Molecular Structure; Neurology; Protein Structure, Secondary; X-Ray Diffraction; Human; Nervous system diseases; Alzheimer disease; Molecular interaction; Glycoproteins; X ray diffraction; Characterization; Molecular assembly; Degenerative disease; Amyloid; Models; Fibril; Crystalline structure
• ISSN: 0006-3495; 1542-0086
• DOI: 10.1016/S0006-3495(93)81393-6

To elucidate the relation between amyloid fibril formation in Alzheimer disease and the primary structure of the beta/A4 protein, which is the major component of the amyloid, we have been investigating the ability of peptides sharing sequences with beta/A4 to form fibrils in vitro. In previous studies we focused on the macroscopic morphology of the assemblies formed by synthetic peptides corresponding in sequence to different regions of this protein. In the present study we analyze the x-ray diffraction patterns obtained from these assemblies. All specimens showed wide angle reflections that could be indexed by an orthogonal lattice of beta-crystallites having unit cell dimensions a = 9.4 A, b = 7 A, and c = 10 A, where a refers to hydrogen bonding direction, b to polypeptide chain direction, and c to intersheet direction. Given the amino acid sequence of beta/A4 as NH2-DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIAT-COOH, we found that, based on their orientation and assembly, the analogues could be classified into three groups: Group A, residues 19–28, 13–28, 12–28, 11–28, 9–28, 1–28, 1–38, 1–40, 6–25, 11–25 and 34–42; Group B, residues 18–28, 17–28, and 15–28; and Group C, residues 22–35 and 26–33. For Groups A and C, the sharpest reflections were (h00), indicating that the assemblies were fibrillar, i.e., elongated in a single direction. Lateral alignment of the crystallites in Group A account for its cross-beta pattern, in which the hydrogen bonding (H-bonding) direction is the fiber (rotation) axis. By comparison, the beta-crystallites of Group C had no preferential orientation, thus giving circular scattering. For Group B, the sharpest reflections were (h0l) on the meridian, indicating that the assemblies were plate-like, i.e., extended in two directions. A series of equatorial Bragg reflections having a 40 A period indicated regular stacking of the plates, and the rotation axis was normal to the surface of the plates. Of the Group A peptides, the analogues 11–28 and 6–25 showed intensity maxima on the equator as well as on higher layer lines, indicating that the beta-crystallites are highly ordered relative to one another in the axial, H-bonding direction. This sampling of the layer lines by a larger period (60 A) suggests that the beta-crystallites are arrayed either in cylindrical or small restricted crystalline lattices.