Formation of native prions from minimal components in vitro
Formation of native prions from minimal components in vitro
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DOI：
10.1073/pnas.0702662104
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发表时间：
2007-06-05
影响因子：
11.1
通讯作者：
Supattapone, Surachai
中科院分区：
文献类型：
--
作者：
Deleault, Nathan R.;Harris, Brent T.;Supattapone, Surachai
宿主蛋白 PrPc 构象变化为疾病相关亚型 PrP5c，似乎在克雅氏病和瘙痒病等朊病毒疾病的发病机制中发挥着关键作用。然而，神经元中产生传染性朊病毒的基本机制仍不清楚。为了从生化角度研究朊病毒形成的机制，我们使用蛋白质错误折叠循环扩增 (PMCA) 技术，使用仅含有天然 PrPc 和共纯化脂质分子的制剂进行了一系列实验。这些实验表明，在纯化系统中成功 PMCA 传播 PrPSc 分子需要辅助聚阴离子分子。此外，我们发现在没有预先存在的朊病毒的情况下，PrPSc 分子可以从这些确定的成分重新形成。将含有朊病毒种子或自发产生的 PrPSc 分子的样品接种到仓鼠体内会引起瘙痒病，这种病在第二次传代时可传播。这些结果表明，能够感染野生型仓鼠的朊病毒可以由最少的一组成分形成，包括天然的 PrPc 分子、共纯化的脂质分子和合成的聚阴离子。
The conformational change of a host protein, PrPc, into a diseaseassociated isoform, PrP5c, appears to play a critical role in the pathogenesis of prion diseases such as Creutzfeldt-Jakob disease and scrapie. However, the fundamental mechanism by which infectious prions are produced in neurons remains unknown. To investigate the mechanism of prion formation biochemically, we conducted a series of experiments using the protein misfolding cyclic amplification (PMCA) technique with a preparation containing only native PrPc and copurified lipid molecules. These experiments showed that successful PMCA propagation of PrPSc molecules in a purified system requires accessory polyanion molecules. In addition, we found that PrPSc molecules could be formed de novo from these defined components in the absence of preexisting prions. Inoculation of samples containing either prion-seeded or spontaneously generated PrPSc molecules into hamsters caused scrapie, which was transmissible on second passage. These results show that prions able to infect wild-type hamsters can be formed from a minimal set of components including native PrPc molecules, copurified lipid molecules, and a synthetic polyanion.