Department of Neurology, David Geffen School of Medicine at UCLA , Neuroscience Research Building, Room 445, 635 Charles E. Young Drive South, Los Angeles, CA 90095-7334, USA.
Biochemistry. 2012 Aug 14;51(32):6342-9. doi: 10.1021/bi3007687. Epub 2012 Aug 3.
The predominant working hypothesis of Alzheimer's disease is that the proximate pathologic agents are oligomers of the amyloid β-protein (Aβ). "Oligomer" is an ill-defined term. Many different types of oligomers have been reported, and they often exist in rapid equilibrium with monomers and higher-order assemblies. This has made formal structure-activity determinations difficult. Recently, Ono et al. [Ono, K., et al. (2009) Proc. Natl. Acad. Sci. U.S.A. 106, 14745-14750] used rapid, zero-length, in situ chemical cross-linking to stabilize the oligomer state, allowing the isolation and study of pure populations of oligomers of a specific order (number of Aβ monomers per assembly). This approach was successful but highly laborious and time-consuming, precluding general application of the method. To overcome these difficulties, we developed a "continuous flow reactor" with the ability to produce theoretically unlimited quantities of chemically stabilized Aβ oligomers. We show, in addition to its utility for Aβ, that this method can be applied to a wide range of other amyloid-forming proteins.
阿尔茨海默病的主要工作假说认为，淀粉样β-蛋白（Aβ）的寡聚体是最接近的致病因子。“寡聚体”是一个定义不明确的术语。已经报道了许多不同类型的寡聚体，它们通常与单体和更高阶的聚集体处于快速平衡状态。这使得正式的结构-活性测定变得困难。最近，Ono 等人[Ono，K.，等人。（2009）Proc。 Natl。Acad。Sci。美国 106，14745-14750]使用快速、零长度、原位化学交联来稳定寡聚体状态，允许分离和研究特定顺序（每个组装体的 Aβ单体数）的纯寡聚体群体。该方法是成功的，但非常繁琐和耗时，排除了该方法的普遍应用。为了克服这些困难，我们开发了一种“连续流反应器”，具有产生理论上无限量化学稳定 Aβ寡聚体的能力。我们不仅证明了它在 Aβ 中的实用性，还证明了该方法可以应用于广泛的其他形成淀粉样蛋白的蛋白质。
