title: Experimentally derived structural constraints for amyloid fibrils of wild-type transthyretin.
abstract: Transthyretin (TTR) is a largely β-sheet serum protein responsible for transporting thyroxine and vitamin A. TTR is found in amyloid deposits of patients with senile systemic amyloidosis. TTR mutants lead to familial amyloidotic polyneuropathy and familial amyloid cardiomyopathy, with an earlier age of onset. Studies of amyloid fibrils of familial amyloidotic polyneuropathy mutant TTR suggest a structure similar to the native state with only a simple opening of a β-strand-loop-strand region exposing the two main β-sheets of the protein for fibril elongation. However, we find that the wild-type TTR sequence forms amyloid fibrils that are considerably different from the previously suggested amyloid structure. Using protease digestion with mass spectrometry, we observe the amyloid core to be primarily composed of the C-terminal region, starting around residue 50. Solid-state NMR measurements prove that TTR differs from other pathological amyloids in not having an in-register parallel β-sheet architecture. We also find that the TTR amyloid is incapable of binding thyroxine as monitored by either isothermal calorimetry or 1,8-anilinonaphthalene sulfonate competition. Taken together, our experiments are consistent with a significantly different configuration of the β-sheets compared to the previously suggested structure.
authors.affiliation: Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda Maryland, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Bateman
authors.forename: Robert
authors.initials: R
authors.lastname: Tycko
authors.forename: Reed B
authors.initials: RB
authors.lastname: Wickner
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000648:ultrastructure; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D011228:Prealbumin / Q000737:chemistry* / Q000648:ultrastructure; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013974:Thyroxine / Q000378:metabolism
chemical_list: D000682:Amyloid; D011228:Prealbumin; D013974:Thyroxine
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