X-ray scattering study of the effect of hydration on the cross-beta structure of amyloid fibrils.
We have investigated the effect of sample hydration on the wide-angle X-ray scattering patterns of amyloid fibrils from two different sources, hen egg white lysozyme (HEWL) and an 11-residue peptide taken from the sequence of transthyretin (TTR105-115). Both samples show an inter-strand reflection at 4.7 A and an inter-sheet reflection which occurs at 8.8 and approximately 10 A for TTR105-115 and HEWL fibrils, respectively. The positions, widths, and relative intensities of these reflections are conserved in patterns obtained from dried stalks and hydrated samples over a range of fibril concentrations. In 2D scattering patterns obtained from flow-aligned hydrated samples, the inter-strand and inter-sheet reflections showed, respectively, axial and equatorial alignment relative to the fibril axis, characteristic of the cross-beta structure. Our results show that the cross-beta structure of the fibrils is not a product of the dehydrating conditions typically employed to produce aligned samples, but is conserved in individual fibrils in hydrated samples under dilute conditions comparable to those associated with other biophysical and spectroscopic techniques. This suggests a structure consisting of a stack of two or more sheets whose interfaces are inaccessible to bulk water.
Squires
Adam M
AM
BSS Sector, Cavendish Laboratory, The University of Cambridge, Madingley Road, Cambridge CB3 0HE, UK. a.m.squires@reading.ac.uk
Devlin
Glyn L
GL
Gras
Sally L
SL
Tickler
Anna K
AK
MacPhee
Cait E
CE
Dobson
Christopher M
CM
D000682:Amyloid / Q000737:chemistry*; D055592:Biophysical Phenomena; D001703:Biophysics; D004527:Egg Proteins / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D011228:Prealbumin / Q000737:chemistry; D017433:Protein Structure, Secondary; D014867:Water / Q000737:chemistry; D014961:X-Ray Diffraction
D000682:Amyloid; D004527:Egg Proteins; D010446:Peptide Fragments; D011228:Prealbumin; D014867:Water; D009113:Muramidase