Abstract
Misfolding of the natively α-helical prion protein into a β-sheet rich isoform is related to various human diseases such as Creutzfeldt-Jakob disease and Gerstmann-Sträussler-Scheinker syndrome. In humans, the disease phenotype is modified by a methionine/valine polymorphism at codon 129 of the prion protein gene. Using a combination of hydrogen/deuterium exchange coupled to NMR spectroscopy, hydroxyl radical probing detected by mass spectrometry, and site-directed mutagenesis, we demonstrate that stop mutants of the human prion protein have a conserved amyloid core. The 129 residue is deeply buried in the amyloid core structure, and its mutation strongly impacts aggregation. Taken together the data support a critical role of the polymorphic residue 129 of the human prion protein in aggregation and disease.
Highlights
In human prion diseases, the phenotype is modified by a methionine/valine polymorphism at codon 129
Using a combination of hydrogen/deuterium exchange coupled to NMR spectroscopy, hydroxyl radical probing detected by mass spectrometry, and site-directed mutagenesis, we demonstrate that stop mutants of the human prion protein have a conserved amyloid core
Using a combination of hydrogen/deuterium (H/D) exchange coupled to NMR spectroscopy, hydroxyl radical probing detected by mass spectrometry, and site-directed mutagenesis, we demonstrate that the valine/methionine residue 129 is deeply buried in the amyloid core of the stop mutants, supporting its critical role in aggregation and disease
Summary
The phenotype is modified by a methionine/valine polymorphism at codon 129. The disease phenotype is modified by a methionine/valine polymorphism at codon 129 of the prion protein gene. Using a combination of hydrogen/deuterium exchange coupled to NMR spectroscopy, hydroxyl radical probing detected by mass spectrometry, and site-directed mutagenesis, we demonstrate that stop mutants of the human prion protein have a conserved amyloid core. Mass spectrometry-coupled exchange experiments of amyloid fibrils of human prion protein (humPrP) comprising residues 90 –231 indicated strong solvent protection in the region encompassing residues ϳ160 –230 [15]. Using a combination of hydrogen/deuterium (H/D) exchange coupled to NMR spectroscopy, hydroxyl radical probing detected by mass spectrometry, and site-directed mutagenesis, we demonstrate that the valine/methionine residue 129 is deeply buried in the amyloid core of the stop mutants, supporting its critical role in aggregation and disease
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