Probing the role of backbone hydrogen bonding in beta-amyloid fibrils with inhibitor peptides containing ester bonds at alternate positions.
Protein-protein interactions are frequently mediated by stable, intermolecular beta-sheets. A number of cytokines and the HIV Protease, for example, dimerize through beta-sheet motifs. Evidence also suggests that the macromolecular assemblies of peptides and proteins in amyloid fibrils are stabilized by intermolecular beta-sheets. In this paper, we report that interfering with the backbone hydrogen bonding of an amyloidgenic peptide (Abeta16-20) by replacing amide bonds with ester bonds prevents the aggregation of the peptide. The ester bonds were incorporated in an alternating fashion so that the peptide presents two unique hydrogen bonding faces when arrayed in an extended, beta-strand conformation; one face of the peptide has normal hydrogen bonding capabilities, but the other face is missing amide protons and its ability to hydrogen bond is severely limited. Analytical ultracentrifugation experiments demonstrate that this ester peptide, Abeta16-20e, is predominantly monomeric under solution conditions, unlike the fibril-forming Abeta16-20 peptide. Abeta16-20e also inhibits the aggregation of the Abeta1-40 peptide and disassembles preformed Abeta1-40 fibrils. These results suggest that backbone hydrogen bonding is critical for the assembly of amyloid fibrils.
Gordon
David J
DJ
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637, USA. djgordon@midway.uchicago.edu
Meredith
Stephen C
SC
D000595:Amino Acid Sequence; D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis / Q000737:chemistry* / Q000648:ultrastructure; D052160:Benzothiazoles; D003224:Congo Red / Q000032:analysis; D003432:Cross-Linking Reagents / Q000032:analysis; D004161:Diphenylhexatriene / Q000032:analysis; D004952:Esters; D005456:Fluorescent Dyes / Q000032:analysis; D006801:Humans; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis / Q000737:chemistry* / Q000648:ultrastructure; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000032:analysis; D011485:Protein Binding; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D021241:Spectrometry, Mass, Electrospray Ionization; D013844:Thiazoles / Q000032:analysis; D014461:Ultracentrifugation
D016229:Amyloid beta-Peptides; D052160:Benzothiazoles; D003432:Cross-Linking Reagents; D004952:Esters; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D013844:Thiazoles; C078592:amyloid beta-protein (1-40); D004161:Diphenylhexatriene; C009462:thioflavin T; D003224:Congo Red; D010649:Phenylalanine; C053764:benzoylphenylalanine