Schütz Anne K, Habenstein Birgit, Luckgei Nina, Bousset Luc, Sourigues Yannick, Nielsen Anders B, Melki Ronald, Böckmann Anja, Meier Beat H
Physical Chemistry, ETH Zürich, Wolfgang-Pauli-Strasse 10, 8093, Zurich, Switzerland.
Biomol NMR Assign. 2014 Oct;8(2):349-56. doi: 10.1007/s12104-013-9515-1. Epub 2013 Aug 14.
Sup35p is a yeast prion and is responsible for the [PSI(+)] trait in Saccharomyces cerevisiae. With 685 amino acids, full-length soluble and fibrillar Sup35p are challenging targets for structural biology as they cannot be investigated by X-ray crystallography or NMR in solution. We present solid-state NMR studies of fibrils formed by the full-length Sup35 protein. We detect an ordered and rigid core of the protein that gives rise to narrow and strong peaks, while large parts of the protein show either static disorder or dynamics on time scales which interfere with dipolar polarization transfer or shorten the coherence lifetime. Thus, only a small subset of resonances is observed in 3D spectra. Here we describe in detail the sequential assignments of the 22 residues for which resonances are observed in 3D spectra: their chemical shifts mostly corresponding to β-sheet secondary structure. We suspect that these residues form the amyloid core of the fibril.
Sup35p是一种酵母朊病毒，负责酿酒酵母中的[PSI(+)]特性。全长可溶性和纤维状的Sup35p含有685个氨基酸，对于结构生物学来说是具有挑战性的研究对象，因为它们无法通过X射线晶体学或溶液中的核磁共振进行研究。我们展示了对全长Sup35蛋白形成的纤维进行的固态核磁共振研究。我们检测到该蛋白有一个有序且刚性的核心，它产生窄而强的峰，而该蛋白的大部分区域要么表现出静态无序，要么在时间尺度上存在动力学，这会干扰偶极极化转移或缩短相干寿命。因此，在三维谱中仅观察到一小部分共振信号。在这里，我们详细描述了在三维谱中观察到共振信号的22个残基的序列归属：它们的化学位移大多对应于β-折叠二级结构。我们怀疑这些残基构成了纤维的淀粉样核心。
