Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH 43210, USA.
Department of Physiology and Biophysics, Case Western Reserve University, Cleveland, OH 44106, USA.
J Struct Biol. 2019 Apr 1;206(1):36-42. doi: 10.1016/j.jsb.2018.04.002. Epub 2018 Apr 18.
The C-terminally truncated Y145Stop variant of prion protein (PrP23-144), which is associated with heritable PrP cerebral amyloid angiopathy in humans and also capable of triggering a transmissible prion disease in mice, serves as a useful in vitro model for investigating the molecular and structural basis of amyloid strains and cross-seeding specificities. Here, we determine the protein-solvent interfaces in human PrP23-144 amyloid fibrils generated from recombinant C,N-enriched protein and incubated in aqueous solution containing paramagnetic Cu(II)-EDTA, by measuring residue-specific N longitudinal paramagnetic relaxation enhancements using two-dimensional magic-angle spinning solid-state NMR spectroscopy. To further probe the interactions of the amyloid core residues with solvent molecules we perform complementary measurements of amide hydrogen/deuterium exchange detected by solid-state NMR and solution NMR methods. The solvent accessibility data are evaluated in the context of the structural model for human PrP23-144 amyloid.
C 端截短的 Y145Stop 突变朊病毒蛋白（PrP23-144）与人类遗传性朊病毒脑淀粉样血管病有关，也能够在小鼠中引发传染性朊病毒病，它是研究淀粉样蛋白株和交叉播种特异性的分子和结构基础的有用体外模型。在这里，我们通过使用二维魔角旋转固态 NMR 光谱测量残基特异性 N 纵向顺磁弛豫增强，来确定从富含 C、N 的重组蛋白生成并在含有顺磁 Cu(II)-EDTA 的水溶液中孵育的人 PrP23-144 淀粉样纤维中的蛋白质-溶剂界面。为了进一步探究淀粉样核心残基与溶剂分子的相互作用，我们通过固态 NMR 和溶液 NMR 方法进行酰胺氢/氘交换检测的互补测量。在人 PrP23-144 淀粉样结构模型的背景下评估溶剂可及性数据。
