Intermolecular structure determination of amyloid fibrils with magic-angle spinning and dynamic nuclear polarization NMR.
Intermolecular structure determination of amyloid fibrils with magic-angle spinning and dynamic nuclear polarization NMR.
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DOI：
10.1021/ja203756x
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发表时间：
2011-09-07
影响因子：
15
通讯作者：
Griffin, Robert G.
中科院分区：
文献类型：
--
作者：
Bayro, Marvin J.;Debelouchina, Galia T.;Eddy, Matthew T.;Birkett, Neil R.;MacPhee, Catherine E.;Rosay, Melanie;Maas, Werner E.;Dobson, Christopher M.;Griffin, Robert G.
我们描述了旨在阐明淀粉样原纤维的链间结构的魔角旋转核磁共振实验。为此引入了三种方法，两种方法基于长程 13C-13C 相关光谱的分析，第三种基于 13C-15N 光谱中分子间相互作用的识别。我们在对 PI3 激酶 (PI3-SH3) 的 86 个残基 SH3 结构域形成的原纤维的研究中表明，有效的 13C-13C 相关光谱显示出共振简并性，从而建立了淀粉样蛋白原纤维中蛋白质的平行、对准排列。此外，可以规避这种简并性以产生直接的分子间约束。 13C-13C 实验由从混合[15N,12C]/[14N,13C]样品获得的15N-13C相关光谱证实，该样品直接量化链间距离。此外，当在 100 K 下通过动态核极化 (DNP) 提供的信号增强来记录光谱时，我们证明光谱中存在的分子间 15N-13C 约束的数量急剧增加（从 23 到 52）。信息内容的增加是由于信号强度的增强以及导致光谱强度损失的动态过程在低温下被淬灭的事实。因此，低温光谱的采集解决了蛋白质 MAS 光谱中经常遇到的问题。实验总共提供了 111 个分子间 13C-13C 和 15N-13C 约束，这些约束确定 PI3-SH3 蛋白链在淀粉样原纤维内以平行、对准的排列方式排列。
We describe magic-angle spinning NMR experiments designed to elucidate the interstrand architecture of amyloid fibrils. Three methods are introduced for this purpose, two being based on the analysis of long-range 13C-13C correlation spectra and a third based on the identification of intermolecular interactions in 13C-15N spectra. We show, in studies of fibrils formed by the 86-residue SH3 domain of PI3 kinase (PI3-SH3), that efficient 13C-13C correlation spectra display a resonance degeneracy that establishes a parallel, in-register alignment of the proteins in the amyloid fibrils. In addition, this degeneracy can be circumvented to yield direct intermolecular constraints. The 13C-13C experiments are corroborated by 15N-13C correlation spectrum obtained from a mixed [15N,12C]/[14N,13C] sample which directly quantifies interstrand distances. Furthermore, when the spectra are recorded with signal enhancement provided by dynamic nuclear polarization (DNP) at 100 K, we demonstrate a dramatic increase (from 23 to 52) in the number of intermolecular 15N-13C constraints present in the spectra. The increase in the information content is due to the enhanced signal intensities and to the fact that dynamic processes, leading to spectral intensity losses, are quenched at low temperatures. Thus, acquisition of low temperature spectra addresses a problem that is frequently encountered in MAS spectra of proteins. In total the experiments provide 111 intermolecular 13C-13C and 15N-13C constraints that establish that the PI3-SH3 protein strands are aligned in a parallel, in-register arrangement within the amyloid fibril.
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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.
DOI：
10.1073/pnas.0812227106
发表时间：
2009-05-12
影响因子：
11.1
作者：
Carullaa, Natalia;Zhou, Min;Dobson, Christopher M.
通讯作者：
Dobson, Christopher M.
影响因子：
4.4
作者：
BENNETT, AE;RIENSTRA, CM;GRIFFIN, RG
通讯作者：
GRIFFIN, RG
DOI：
10.1073/pnas.252625999
发表时间：
2002-12-24
影响因子：
11.1
作者：
Jaroniec, CP;MacPhee, CE;Griffin, RG
通讯作者：
Griffin, RG