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利用类朊病毒蛋白RepA-WH1解决细菌中的细胞内淀粉样变性问题。

Addressing Intracellular Amyloidosis in Bacteria with RepA-WH1, a Prion-Like Protein.

作者信息

Molina-García Laura, Gasset-Rosa Fátima, Álamo María Moreno-Del, de la Espina Susana Moreno-Díaz, Giraldo Rafael

机构信息

Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas (CSIC), Madrid, Spain.

Department of Cell and Developmental Biology, University College London, London, UK.

出版信息

Methods Mol Biol. 2018;1779:289-312. doi: 10.1007/978-1-4939-7816-8_18.

Abstract

Bacteria are the simplest cellular model in which amyloidosis has been addressed. It is well documented that bacterial consortia (biofilms) assemble their extracellular matrix on an amyloid scaffold, yet very few intracellular amyloids are known in bacteria. Here, we describe the methods we have resorted to characterize in Escherichia coli cells the amyloidogenesis, propagation, and dynamics of the RepA-WH1 prionoid. This prion-like protein, a manifold domain from the plasmid replication protein RepA, itself capable of assembling a functional amyloid, causes when expressed in E. coli a synthetic amyloid proteinopathy, the first model for an amyloid disease with a purely bacterial origin. These protocols are useful to study other intracellular amyloids in bacteria.

摘要

细菌是研究淀粉样变性的最简单细胞模型。有充分的文献记载,细菌群落(生物膜)在淀粉样支架上组装其细胞外基质,但细菌中已知的细胞内淀粉样蛋白却很少。在这里,我们描述了我们用来在大肠杆菌细胞中表征RepA-WH1类朊病毒的淀粉样蛋白生成、传播和动力学的方法。这种类朊病毒蛋白是质粒复制蛋白RepA的一个多结构域,本身能够组装功能性淀粉样蛋白,当在大肠杆菌中表达时会导致一种合成性淀粉样蛋白病,这是第一个具有纯细菌起源的淀粉样疾病模型。这些方案对于研究细菌中的其他细胞内淀粉样蛋白很有用。

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