Comparison of the level, distribution and form of disease-associated prion protein in variant and sporadic Creutzfeldt-Jakob diseased brain using conformation-dependent immunoassay and Western blot
Comparison of the level, distribution and form of disease-associated prion protein in variant and sporadic Creutzfeldt-Jakob diseased brain using conformation-dependent immunoassay and Western blot
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DOI：
10.1099/vir.0.026948-0
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发表时间：
2011-03-01
影响因子：
3.8
通讯作者：
Head, Mark W.
中科院分区：
文献类型：
--
作者：
Choi, Young Pyo;Groener, Albrecht;Head, Mark W.
疾病相关朊病毒蛋白 (PrPSc) 可以通过构象依赖性免疫测定 (CDI) 与细胞亚型 (PrPC) 区分开来。该技术利用了 PrPC 中存在的表位，但只有在 PrPSc 中变性时才能暴露出来。在本研究中，我们利用 CDI 研究了变异型和散发性克雅氏病 (CJD) 不同脑区的 PrPSc，并将结果与使用更常用的蛋白酶消化和蛋白质印迹法获得的结果直接进行比较。总的来说，结果之间有很好的一致性，尽管考虑到不同克雅氏病病例的区域分布，相对丰度存在一定差异。结果在很大程度上证实了先前描述的变异型和散发型克雅氏病针对不同大脑区域的研究。此外，蛋白酶消化和 CDI 检测相结合，首次证明变异型 CJD 大脑中存在 PrPSc，且在标准条件下易发生蛋白水解。
Disease-associated prion protein (PrPSc) can be distinguished from the cellular isoform (PrPC) by conformation-dependent immunoassay (CDI). This technique exploits the presence of an epitope, accessible in PrPC, but only unmasked by denaturation in PrPSc. In this study, we investigated PrPSc in different brain regions in variant and sporadic Creutzfeldt-Jakob disease (CJD) by using CDI, and directly compared the results with those obtained using the more commonly employed protease digestion and Western blotting. In general, there was good agreement between the results, although there were certain discrepancies in relative abundance when the regional distribution in variant CJD cases was considered. The results largely confirmed the previously described targeting of different brain regions by variant and sporadic CJD. Additionally, the combination of protease digestion and CDI detection demonstrated, for the first time, the presence of PrPSc in variant CJD brains that is susceptible to proteolysis under standard conditions.