Sensitivity-Enhanced Solid-State NMR Detection of Structural Differences and Unique Polymorphs in Pico- to Nanomolar Amounts of Brain-Derived and Synthetic 42-Residue Amyloid-β Fibrils.
Sensitivity-Enhanced Solid-State NMR Detection of Structural Differences and Unique Polymorphs in Pico- to Nanomolar Amounts of Brain-Derived and Synthetic 42-Residue Amyloid-β Fibrils.
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DOI：
10.1021/jacs.1c03346
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发表时间：
2021-08-04
影响因子：
15
通讯作者：
Ishii, Yoshitaka
中科院分区：
文献类型：
--
作者：
Wickramasinghe, Ayesha;Xiao, Yiling;Kobayashi, Naohiro;Wang, Songlin;Scherpelz, Kathryn P.;Yamazaki, Toshio;Meredith, Stephen C.;Ishii, Yoshitaka
神经炎斑中的β淀粉样蛋白（Aβ）纤维是阿尔茨海默病（AD）的标志。由于42个残基的Aβ（Aβ42）原纤维是不同Aβ物种中致病性最强的，因此其结构特征对于我们了解AD至关重要。虽然已经报道了Aβ40的几种多晶型物，但先前对在中性pH下制备的Aβ42原纤维的研究基本上只检测到一种结构，具有S形β折叠排列[例如，Xiao等人，天然结构分子2015，22，499]。在此，我们证明了在超快魔角旋转（UFMAS）下通过灵敏度增强的1H检测固态NMR（SSNMR）表征痕量脑源性和合成淀粉样蛋白原纤维结构的可行性。通过利用该技术的高灵敏度，我们首次证明了其适用于高通量筛选痕量的选择性13 C和15 N标记的Aβ42原纤维，该原纤维用约0.01%的患者源性淀粉样蛋白（约10%）制备。4 pmol）作为种子。2D 13 C/1H SSNMR数据的比较显示，在不到10 min的时间内，AD源性Aβ42（~40 nmol或~200 μg）与合成原纤维之间存在显著的结构差异，证实了在~2.5 h内评估约1 pmol脑淀粉样蛋白种子的原纤维结构的可行性。我们还通过1H检测的3D和4D SSNMR首次对合成的完全质子化的Aβ42原纤维进行了结构表征。在自动分配的辅助下，在1H检测方法中完成了痕量（~42 nmol）完全质子化淀粉样蛋白原纤维的主链共振分配。结果表明，这种Aβ42原纤维表现出一种新的折叠或多晶型结构。
Amyloid-β (Aβ) fibrils in neuritic plaques are a hallmark of Alzheimer’s disease (AD). Since the 42-residue Aβ (Aβ42) fibril is the most pathogenic among different Aβ species, its structural characterization is crucial to our understanding of AD. While several polymorphs have been reported for Aβ40, previous studies of Aβ42 fibrils prepared at neutral pH detected essentially only one structure, with an S-shaped β-sheet arrangement [e.g., Xiao et al., Nat. Struct. Mol. Biol. 2015, 22, 499]. Herein, we demonstrate the feasibility of characterizing the structure of trace amounts of brain-derived and synthetic amyloid fibrils by sensitivity-enhanced 1H-detected solid-state NMR (SSNMR) under ultra-fast magic angle spinning (UFMAS). By taking advantage of the high sensitivity of this technique, we first demonstrate its applicability for the high-throughput screening of trace amounts of selectively 13C- and 15N-labeld Aβ42 fibril prepared with ~0.01% patient-derived amyloid (ca. 4 pmol) as a seed. The comparison of 2D 13C/1H SSNMR data revealed marked structural differences between AD-derived Aβ42 (~40 nmol or ~200 μg) and synthetic fibrils in less than 10 min, confirming the feasibility of assessing the fibril structure from ~1 pmol of brain amyloid seed in ~2.5 h. We also present the first structural characterization of synthetic fully-protonated Aβ42 fibril by 1H-detected 3D and 4D SSNMR. With procedures assisted by automated assignments, main-chain resonance assignments were completed for trace amounts (~42 nmol) of a fully-protonated amyloid fibril in the 1H-detection approach. The results suggest that this Aβ42 fibril exhibits a novel fold or polymorph structure.
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影响因子：
15
作者：
Colvin MT;Silvers R;Frohm B;Su Y;Linse S;Griffin RG
通讯作者：
Griffin RG
DOI：
10.1126/science.aao2825
发表时间：
2017-10-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gremer L;Schölzel D;Schenk C;Reinartz E;Labahn J;Ravelli RBG;Tusche M;Lopez-Iglesias C;Hoyer W;Heise H;Willbold D;Schröder GF
通讯作者：
Schröder GF
影响因子：
2.2
作者：
Ishii, Y;Tycko, R
通讯作者：
Tycko, R
影响因子：
2.9
作者：
Burdick, D;Kosmoski, J;Glabe, CG
通讯作者：
Glabe, CG
影响因子：
4.9
作者：
Ghosh, Ujjayini;Yau, Wai-Ming;Tycko, Robert
通讯作者：
Tycko, Robert