全长重组朊蛋白的纯化与纤维化
Purification and fibrillation of full-length recombinant PrP.
作者信息
Makarava Natallia, Baskakov Ilia V
机构信息
Center for Biomedical Engineering and Technology, Department of Anatomy and Neurobiology, University of Maryland, Baltimore, MD, USA.
出版信息
Methods Mol Biol. 2012;849:33-52. doi: 10.1007/978-1-61779-551-0_4.
Misfolding and aggregation of prion protein (PrP) is related to several neurodegenerative diseases in humans such as Creutzfeldt-Jacob disease, fatal familial insomnia, and Gerstmann-Straussler-Sheinker disease. Certain applications in prion area require recombinant PrP of high purity and quality. Here, we report an experimental procedure for expression and purification of full-length mammalian PrP. This protocol has been proved to yield PrP of extremely high purity that lacks PrP adducts, which are normally generated as a result of spontaneous oxidation or degradation. We also describe methods for the preparation of amyloid fibrils from recombinant PrP in vitro. Recombinant PrP fibrils can be used as a noninfectious synthetic surrogate of PrP(Sc) for development of prion diagnostics including the generation of PrP(Sc)-specific antibody.
朊病毒蛋白（PrP）的错误折叠和聚集与人类的几种神经退行性疾病有关，如克雅氏病、致死性家族性失眠症和格斯特曼-施特劳斯勒-谢克尔病。朊病毒领域的某些应用需要高纯度和高质量的重组PrP。在此，我们报告了一种全长哺乳动物PrP表达和纯化的实验方法。该方案已被证明能产生极高纯度的PrP，且不含通常因自发氧化或降解而产生的PrP加合物。我们还描述了体外从重组PrP制备淀粉样纤维的方法。重组PrP纤维可作为PrP(Sc)的非感染性合成替代物，用于朊病毒诊断的开发，包括产生PrP(Sc)特异性抗体。
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