Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization.
Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization.
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作者：
Ryan TM;Caine J;Mertens HD;Kirby N;Nigro J;Breheney K;Waddington LJ;Streltsov VA;Curtain C;Masters CL;Roberts BR
阿尔茨海默病是老年人痴呆症的主要原因。病理上表现为淀粉样斑块的存在和患者脑组织内神经元的丧失。现在普遍假设纤维状结构代表一种惰性结构。试图表征Aβ纤维和中间寡聚结构形成的生物物理和毒性分析通常涉及制备主要为单体的样品；实现这一目标的最常用方法是使用含氟有机溶剂1,1,1,3,3,3-六氟-2-丙醇（HFIP）。最近的证据表明，这种方法在产生无骨料溶液时不是100%有效。我们使用动态光散射、尺寸排除色谱和小角度x射线散射表明，HFIP预处理的Aβ肽溶液确实是这种情况，显示出低聚物和聚集物质的比例增加，并且聚集倾向增加。此外，我们表明，一种替代技术，涉及处理强碱的结果更均匀的溶液，大部分是单体。这些溶解和控制Aβ寡聚物状态的技术是未来生物物理和毒性分析的有价值的起点。
Alzheimer’s disease is the leading cause of dementia in the elderly. Pathologically it is characterized by the presence of amyloid plaques and neuronal loss within the brain tissue of affected individuals. It is now widely hypothesised that fibrillar structures represent an inert structure. Biophysical and toxicity assays attempting to characterize the formation of both the fibrillar and the intermediate oligomeric structures of Aβ typically involves preparing samples which are largely monomeric; the most common method by which this is achieved is to use the fluorinated organic solvent 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). Recent evidence has suggested that this method is not 100% effective in producing an aggregate free solution. We show, using dynamic light scattering, size exclusion chromatography and small angle X-ray scattering that this is indeed the case, with HFIP pretreated Aβ peptide solutions displaying an increased proportion of oligomeric and aggregated material and an increased propensity to aggregate. Furthermore we show that an alternative technique, involving treatment with strong alkali results in a much more homogenous solution that is largely monomeric. These techniques for solubilising and controlling the oligomeric state of Aβ are valuable starting points for future biophysical and toxicity assays.
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DOI：
10.1016/s0006-291x(84)80190-4
发表时间：
1984-01-01
影响因子：
3.1
作者：
GLENNER, GG;WONG, CW
通讯作者：
WONG, CW
影响因子：
6.1
作者：
Konarev, PV;Volkov, VV;Svergun, DI
通讯作者：
Svergun, DI
DOI：
10.1016/s0006-291x(02)00346-7
发表时间：
2002-05-17
影响因子：
3.1
作者：
Blanchard, BJ;Thomas, VL;Ingram, VM
通讯作者：
Ingram, VM
影响因子：
3
作者：
Huang, XD;Atwood, CS;Bush, AI
通讯作者：
Bush, AI
影响因子：
4.2
作者：
Ciccotosto, Giuseppe D.;Tew, Deborah J.;Cappai, Roberto
通讯作者：
Cappai, Roberto