SEPARATION AND PROPERTIES OF CELLULAR AND SCRAPIE PRION PROTEINS
SEPARATION AND PROPERTIES OF CELLULAR AND SCRAPIE PRION PROTEINS
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DOI：
10.1073/pnas.83.8.2310
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发表时间：
1986-04-01
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：
MEYER, RK;MCKINLEY, MP;PRUSINER, SB
羊瘙痒症朊病毒的纯化制剂含有Mr为27，000 - 30，000的唾液酸糖蛋白，命名为PrP 27 - 30，其通过有限的蛋白水解从羊瘙痒症朊病毒蛋白[Mr，33，000 - 35，000（PrP 33 - 35Sc）]衍生而来。在这些相同的蛋白水解条件下，相同大小的细胞蛋白（PrP 33 - 35C）被完全降解。仓鼠脑的亚细胞分级显示PrP33 - 35Sc和PrP33 - 35C仅见于膜组分。NaCl、EDTA和渗透压休克未能从微粒体膜中释放朊病毒蛋白。这些微粒体组分的电子显微镜显示膜囊泡，但不是朊病毒淀粉样蛋白棒。洗涤剂处理羊瘙痒病感染的膜溶解PrP 33 - 35 C，而PrPSc聚集成淀粉样蛋白棒; PrP 33 - 35 C的浓度与从未感染的对照脑制备的类似组分中回收的浓度相似。PrP 33 - 35Sc的明显的两亲性特征可以解释羊瘙痒病感染性与膜和淀粉样蛋白丝的关联。
Purified preparations of scrapie prions contain a sialoglycoprotein of Mr 27,000-30,000, designated PrP 27-30, which is derived from the scrapie prion protein [Mr, 33,000-35,000 (PrP 33-35Sc)] by limited proteolysis. Under these same conditions of proteolysis, a cellular protein of the same size (PrP 33-35C) is completely degraded. Subcellular fractionation of hamster brain showed that both PrP 33-35Sc and PrP 33-35C were found only in membrane fractions. NaCl, EDTA, and osmotic shock failed to release the prion proteins from microsomal membranes. Electron microscopy of these microsomal fractions showed membrane vesicles but not prion amyloid rods. Detergent treatment of scrapie-infected membranes solubilized PrP 33-35C, while PrPSc aggregated into amyloid rods; the concentration of PrP 33-35C was similar to that recovered from analogous fractions prepared from uninfected control brains. The apparent amphipathic character of the PrP 33-35Sc may explain the association of scrapie infectivity with both membranes and amyloid filaments.