Continuous Flow Reactor for the Production of Stable Amyloid Protein Oligomers

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• 作者：Eric Yale Hayden ; David B. Teplow
• 刊名：Biochemistry
• 出版年：2012
• 出版时间：August 14, 2012
• 年：2012
• 卷：51
• 期：32
• 页码：6342-6349
• 全文大小：398K
• 年卷期：v.51,no.32(August 14, 2012)
• ISSN：1520-4995

文摘

The predominant working hypothesis of Alzheimer鈥檚 disease is that the proximate pathologic agents are oligomers of the amyloid 尾-protein (A尾). 鈥淥ligomer鈥?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鈥揳ctivity determinations difficult. Recently, Ono et al. [Ono, K., et al. (2009) Proc. Natl. Acad. Sci. U.S.A. 106, 14745鈥?4750] 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 A尾 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 鈥渃ontinuous flow reactor鈥?with the ability to produce theoretically unlimited quantities of chemically stabilized A尾 oligomers. We show, in addition to its utility for A尾, that this method can be applied to a wide range of other amyloid-forming proteins.