title: The organization of aromatic side groups in an amyloid fibril probed by solid-state 2H and 19F NMR spectroscopy.
abstract: Some 25 diseases are associated with proteins and peptides that assemble into amyloid fibrils composed of beta-strands connected by hydrogen bonds oriented parallel to the fiber long axis. There is mounting evidence that amyloid formation involves specific interactions between amino acid side groups, which bring together beta-sheets to form layers with buried and exposed faces. This work demonstrates how a combination of solid-state 2H and 19F NMR experiments can provide constraints on fibril architecture by probing the environment and spatial organisation of aromatic side groups. It is shown that phenylalanine rings within fibrils formed by a decapeptide fragment of the islet amyloid polypeptide, amylin, are highly motionally restrained and are situated within 6.5 A of one another. Taken together with existing structural constraints for this peptide, these results are consistent with a fibril architecture that comprises layers of two or more beta-sheets, with the aromatic residues facing into the inter-sheet space and possibly engaged in pi-pi interactions. The methods presented will be of general utility in exploring the architecture of fibrils of larger, full-length peptides and proteins, including amylin itself.
authors.affiliation: Faculty of Life Sciences, University of Manchester, P.O. Box 88, Manchester M60 1QD, United Kingdom.
authors.forename: Edward
authors.initials: E
authors.lastname: Jack
authors.forename: Matthew
authors.initials: M
authors.lastname: Newsome
authors.forename: Peter G
authors.initials: PG
authors.lastname: Stockley
authors.forename: Sheena E
authors.initials: SE
authors.lastname: Radford
authors.forename: David A
authors.initials: DA
authors.lastname: Middleton
mesh_terms: D000682:Amyloid / Q000737:chemistry*; D003903:Deuterium / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry; D058228:Islet Amyloid Polypeptide; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015394:Molecular Structure; D010446:Peptide Fragments / Q000737:chemistry*; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D017433:Protein Structure, Secondary
chemical_list: D000682:Amyloid; C521761:IAPP(20-29) peptide; D058228:Islet Amyloid Polypeptide; D010446:Peptide Fragments; D005461:Fluorine; D010649:Phenylalanine; D003903:Deuterium