Amyloid fibril-like structure underlies the aggregate structure across the pH range for beta-lactoglobulin.
The protein beta-lactoglobulin aggregates into two apparently distinct forms under different conditions: amyloid fibrils at pH values away from the isoelectric point, and spherical aggregates near it. To understand this apparent dichotomy in behavior, we studied the internal structure of the spherical aggregates by employing a range of biophysical approaches. Fourier transform infrared studies show the aggregates have a high beta-sheet content that is distinct from the native beta-lactoglobulin structure. The structures also bind the amyloidophilic dye thioflavin-T, and wide-angle x-ray diffraction showed reflections corresponding to spacings typically observed for amyloid fibrils composed of beta-lactoglobulin. Combined with small-angle x-ray scattering data indicating the presence of one-dimensional linear aggregates at the molecular level, these findings indicate strongly that the aggregates contain amyloid-like substructure. Incubation of beta-lactoglobulin at pH values increasingly removed from the isoelectric point resulted in the increasing appearance of fibrillar species, rather than spherical species shown by electron microscopy. Taken together, these results suggest that amyloid-like beta-sheet structures underlie protein aggregation over a much broader range of conditions than previously believed. Furthermore, the results suggest that there is a continuum of beta-sheet structure of varying regularity underlying the aggregate morphology, from very regular amyloid fibrils at high charge to short stretches of amyloid-like fibrils that associate together randomly to form spherical particles at low net charge.
Krebs
Mark R H
MR
Cavendish Laboratory, University of Cambridge, Cambridge, United Kingdom.
Devlin
Glyn L
GL
Donald
Athene M
AM
D000682:Amyloid / Q000737:chemistry / Q000648:ultrastructure*; D000818:Animals; D052160:Benzothiazoles; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry / Q000648:ultrastructure*; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D017550:Spectroscopy, Fourier Transform Infrared; D013844:Thiazoles; D014961:X-Ray Diffraction
D000682:Amyloid; D052160:Benzothiazoles; D007782:Lactoglobulins; D013844:Thiazoles; C009462:thioflavin T