SCRAPIE-INFECTED MURINE NEURO-BLASTOMA CELLS PRODUCE PROTEASE-RESISTANT PRION PROTEINS
SCRAPIE-INFECTED MURINE NEURO-BLASTOMA CELLS PRODUCE PROTEASE-RESISTANT PRION PROTEINS
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DOI：
10.1128/jvi.62.5.1558-1564.1988
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发表时间：
1988-05-01
影响因子：
5.4
通讯作者：
PRUSINER, SB
中科院分区：
文献类型：
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作者：
BUTLER, DA;SCOTT, MRD;PRUSINER, SB
痒病和克雅氏病是由朊病毒引起的传染性、退行性神经系统疾病。相当多的证据表明朊病毒含有抗蛋白酶的唾液酸糖蛋白，称为PrPSc，由细胞基因编码。朊病毒蛋白（PrP）基因也编码称为PrPC的正常细胞蛋白。我们建立了支持小鼠瘙痒病或克雅氏病朊病毒复制的克隆细胞系。小鼠神经母细胞瘤N2 a细胞暴露于小鼠瘙痒症朊病毒，随后克隆。有限的蛋白酶K消化后，三个PrP-免疫反应性蛋白质的表观分子量范围在20和30千道尔顿之间的羊瘙痒病感染的N2 a细胞的提取物中检测到蛋白质（免疫）印迹。这些PrPSc分子的真实性是通过使用针对与朊病毒蛋白的一部分相对应的合成肽产生的单特异性抗血清来建立的。这些克隆合成PrPSc分子具有痒病朊病毒感染性的生物测定法测定，克隆没有PrPSc未能证明感染性。在羊瘙痒病感染的N2 a细胞中检测到PrPSc分子支持了PrPSc是传染性羊瘙痒病颗粒的组分的论点，并为朊病毒疾病的研究开辟了新的途径。
Scrapie and Creutzfeldt-Jakob disease are transmissible, degenerative neurological diseases caused by prions. Considerable evidence argues that prions contain protease-resistant sialoglycoproteins, designated PrPSc, encoded by a cellular gene. The prion protein (PrP) gene also encodes a normal cellular protein designated PrPC. We established clonal cell lines which support the replication of mouse scrapie or Creutzfeldt-Jakob disease prions. Mouse neuroblastoma N2a cells were exposed to mouse scrapie prions and subsequently cloned. After limited proteinase K digestion, three PrP-immunoreactive proteins with apparent molecular masses ranging between 20 and 30 kilodaltons were detected in extracts of scrapie-infected N2a cells by Western (immono-) blotting. The authenticity of these PrPSc molecules was established by using monospecific antiserum raised against a synthetic peptide corresponding to a portion of the prion protein. Those clones synthesizing PrPSc molecules possessed scrapie prion infectivity as measured by bioassay; clones without PrPSc failed to demonstrate infectivity. Detection of PrPSc molecules in scrapie-infected N2a cells supports the contention that PrPSc is a component of the infectious scrapie particle and opens new approaches to the study of prion diseases.