IMMUNOAFFINITY PURIFICATION AND NEUTRALIZATION OF SCRAPIE PRION INFECTIVITY
IMMUNOAFFINITY PURIFICATION AND NEUTRALIZATION OF SCRAPIE PRION INFECTIVITY
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DOI：
10.1073/pnas.85.18.6617
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发表时间：
1988-09-01
影响因子：
11.1
通讯作者：
PRUSINER, SB
中科院分区：
文献类型：
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作者：
GABIZON, R;MCKINLEY, MP;PRUSINER, SB
朊病毒是引起绵羊瘙痒症和人类克雅氏病的常见病原体。生物化学和遗传学研究认为，羊瘙痒症朊病毒蛋白（PrPSc）的同种型是朊病毒的主要成分。PrPSc的有限蛋白酶K消化产生27-30 kDa的蛋白质。从羊瘙痒病感染的仓鼠中分离的脑微粒体分散到洗涤剂-脂质-蛋白复合物中后，用27-30 kKa的羊瘙痒病朊蛋白单克隆抗体亲和柱获得PrPSc和羊瘙痒病感染性的共纯化。富集PrPSc。5700-倍，而羊瘙痒症朊病毒感染性被富集。四千倍。朊病毒滴度与PrPSc的比率在整个纯化过程中保持恒定。异源单克隆抗体柱未能结合PrPSc或羊瘙痒病感染性。针对NaDodSO 4/PAGE纯化的27-30 kDa羊瘙痒症朊病毒蛋白的多克隆兔朊病毒蛋白抗血清使分散到去污剂-脂质-蛋白复合物中的羊瘙痒症感染性降低100倍。这些结果代表了直接的免疫学和色谱演示之间的关系PrPSc和朊病毒的感染性，以及提供额外的支持的论点，PrPSc是一个主要组成部分的传染性瘙痒病颗粒。PrPSc是一种宿主编码的蛋白质，这是朊病毒区别于病毒的一个重要特征。
Prions are usnusual infectious pathogens causing scrapie of sheep goats as well as Creutzfeldt-Jakob disease of humans. Biochemical and genetic studies contend that the scrapie isoform of the prion protein (PrPSc) is a major component of the prion. Limited proteinase K digestion of PrPSc produced a protein of 27-30 kDa. After dispersion of brain microsomes isolated from scrapie-infected hamsters into detergent-lipid-protein complexes, copurification of PrPSc and scrapie infectivity was obtained with scrapie prion protein of 27-30 kKa monoclonal antibody-affinity columns. PrPSc was enriched .apprxeq. 5700-fold with respect to total brain protein, whereas scrapie prion infectivity was enriched .apprxeq. 4000-fold. The ratio of prion titer to PrPSc remained constant throughout purification. Heterologous monoclonal antibody columns failed to bind either PrPSc or scrapie infectivity. Polyclonal rabbit prion protein antiserum raised against NaDodSO4/PAGE-purified scrapie prion protein of 27-30 kDa reduced scrapie infectivity dispersed into detergent-lipid-protein complexes by a factor of 100. These results represent direct immunologic and chromatographic demonstrations of a relationship between PrPSc and prion infectivity as well as providing additional support for the contention that PrPSc is a major component of the infectious scrapie particle. That PrPSc is a host-encoded protein is an important feature distinguishing prions from viruses.