Higher order amyloid fibril structure by MAS NMR and DNP spectroscopy.
Higher order amyloid fibril structure by MAS NMR and DNP spectroscopy.
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DOI：
10.1021/ja409050a
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发表时间：
2013-12-26
影响因子：
15
通讯作者：
Griffin RG
中科院分区：
文献类型：
--
作者：
Debelouchina GT;Bayro MJ;Fitzpatrick AW;Ladizhansky V;Colvin MT;Caporini MA;Jaroniec CP;Bajaj VS;Rosay M;Macphee CE;Vendruscolo M;Maas WE;Dobson CM;Griffin RG
蛋白魔角旋转（MAS）NMR光谱已经产生了几个淀粉样蛋白原纤维系统的结构模型，从而提供了有价值的信息，赋予非凡的稳定性的淀粉样蛋白结构的力量和相互作用。然而，尽管取得了这些进展，获得原子分辨率信息描述更高层次的结构组织内的原纤维仍然是一个重大的挑战。在这里，我们详细介绍了MAS NMR实验和样品标记方案，专门设计来探测这种高阶淀粉样蛋白结构，我们已经将它们应用于由淀粉样蛋白转甲状腺素蛋白（TTR（105-115））的11个残基片段形成的原纤维。这些实验使我们不仅能够明确地定义肽β链到β折叠的排列，而且还可以明确地定义每个原丝内的β折叠界面，此外还可以确定导致完整原纤维形成的原丝与原丝接触的性质。我们的努力已经导致了111定量的距离和扭转角的限制（10个残基），描述了不同层次的结构组织。这些实验广泛受益于动态核极化（DNP）的使用，在某些情况下，它使我们能够缩短数据采集时间从几天到几个小时，并显着提高光谱的信噪比。此处鉴定的β-折叠界面和原丝相互作用揭示了结构的局部变化，其导致肽的暴露的N-和C-末端的多个峰以及埋藏在原纤维内部的侧链的不均匀线增宽。
Protein magic angle spinning (MAS) NMR spectroscopy has generated structural models of several amyloid fibril systems, thus providing valuable information regarding the forces and interactions that confer the extraordinary stability of the amyloid architecture. Despite these advances, however, obtaining atomic resolution information describing the higher levels of structural organization within the fibrils remains a significant challenge. Here, we detail MAS NMR experiments and sample labeling schemes designed specifically to probe such higher order amyloid structure and we have applied them to the fibrils formed by an eleven-residue segment of the amyloidogenic protein transthyretin (TTR(105-115)). These experiments have allowed us to define unambiguously not only the arrangement of the peptide β-strands into β-sheets but also the β-sheet interfaces within each protofilament, and in addition to identify the nature of the protofilament-to-protofilament contacts that lead to the formation of the complete fibril. Our efforts have resulted in 111 quantitative distance and torsion angle restraints (10 per residue) that describe the various levels of structure organization. The experiments benefited extensively from the use of dynamic nuclear polarization (DNP), which in some cases allowed us to shorten the data acquisition time from days to hours and to improve significantly the signal-to-noise ratios of the spectra. The β-sheet interface and protofilament interactions identified here revealed local variations in the structure that result in multiple peaks for the exposed N- and C-termini of the peptide and in inhomogeneous line-broadening for the side-chains buried within the interior of the fibrils.
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影响因子：
2.9
作者：
Bayro, Marvin J.;Maly, Thorsten;Birkett, Neil R.;MacPhee, Cait E.;Dobson, Christopher M.;Griffin, Robert G.
通讯作者：
Griffin, Robert G.
影响因子：
3.3
作者：
Caporini, Marc A.;Bajaj, Vikram S.;Veshtort, Mikhail;Fitzpatrick, Anthony;MacPhee, Cait E.;Vendruscolo, Michele;Dobson, Christopher M.;Griffin, Robert G.
通讯作者：
Griffin, Robert G.
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
影响因子：
1.7
作者：
Baldus, M;Petkova, AT;Griffin, RG
通讯作者：
Griffin, RG
影响因子：
2.9
作者：
Andreas, Loren B.;Barnes, Alexander B.;Griffin, Robert G.
通讯作者：
Griffin, Robert G.