Cyclic Amplification of Prion Protein Misfolding
Cyclic Amplification of Prion Protein Misfolding
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DOI：
10.1007/978-1-61779-551-0_14
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Soto, Claudio
中科院分区：
文献类型：
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作者：
Barria, Marcelo A.;Gonzalez-Romero, Dennisse;Soto, Claudio
蛋白质错误折叠循环扩增（PMCA）是一种利用成核依赖性朊病毒复制过程加速PrPC在试管中转化为PrPSc的技术。PMCA使用超声波将PrPSc聚合物片段化，增加感染样品中存在的种子量，而不影响它们作为转化核的能力。在过去的5年里，PMCA已经成为研究朊病毒各个方面的宝贵技术。PMCA技术已被多个研究小组用于了解朊病毒复制的分子机制、朊病毒繁殖中涉及的细胞因子、朊病毒菌株和物种屏障的有趣现象、检测组织和生物体液中的PrPSc以及筛选朊病毒复制的抑制剂。在本章中，我们描述了一个详细的协议的PMCA技术，突出了一些重要的技术方面，以获得一个成功的和可重复的应用的技术。
Protein misfolding cyclic amplification (PMCA) is a technique that takes advantage of the nucleation-dependent prion replication process to accelerate the conversion of PrPC into PrPSc in the test tube. PMCA uses ultrasound waves to fragment the PrPSc polymers, increasing the amount of seeds present in the infected sample without affecting their ability to act as conversion nuclei. Over the past 5 years, PMCA has become an invaluable technique to study diverse aspects of prions. The PMCA technology has been used by several groups to understand the molecular mechanism of prion replication, the cellular factors involved in prion propagation, the intriguing phenomena of prion strains and species barriers, to detect PrPSc in tissues and biological fluids, and to screen for inhibitors against prion replication. In this chapter, we describe a detailed protocol of the PMCA technique, highlighting some of the important technical aspects to obtain a successful and reproducible application of the technology.