Physical Chemistry, ETH Zürich, Wolfgang-Pauli-Strasse 10, CH-8093 Zurich, Switzerland.
J Am Chem Soc. 2010 Oct 6;132(39):13765-75. doi: 10.1021/ja104213j.
We present a strategy to solve the high-resolution structure of amyloid fibrils by solid-state NMR and use it to determine the atomic-resolution structure of the prion domain of the fungal prion HET-s in its amyloid form. On the basis of 134 unambiguous distance restraints, we recently showed that HET-s(218-289) in its fibrillar state forms a left-handed β-solenoid, and an atomic-resolution NMR structure of the triangular core was determined from unambiguous restraints only. In this paper, we go considerably further and present a comprehensive protocol using six differently labeled samples, a collection of optimized solid-state NMR experiments, and adapted structure calculation protocols. The high-resolution structure obtained includes the less ordered but biologically important C-terminal part and improves the overall accuracy by including a large number of ambiguous distance restraints.
我们提出了一种通过固态 NMR 解决淀粉样纤维高分辨率结构的策略，并将其用于确定真菌朊病毒 HET-s 的朊病毒结构域在其淀粉样形式下的原子分辨率结构。基于 134 个明确的距离约束，我们最近表明，其纤维状状态下的 HET-s（218-289）形成左手β-螺线管，并且仅从明确的约束中确定了三角形核心的原子分辨率 NMR 结构。在本文中，我们更进一步，提出了一个全面的方案，该方案使用六个不同标记的样品、一组优化的固态 NMR 实验和改编的结构计算方案。所获得的高分辨率结构包括较无序但具有生物学重要性的 C 末端部分，并通过包含大量模糊距离约束来提高整体准确性。
