PARTIAL-PURIFICATION AND EVIDENCE FOR MULTIPLE MOLECULAR-FORMS OF SCRAPIE AGENT
PARTIAL-PURIFICATION AND EVIDENCE FOR MULTIPLE MOLECULAR-FORMS OF SCRAPIE AGENT
复制标题
DOI：
10.1021/bi00616a021
复制
发表时间：
1978-01-01
期刊：
影响因子：
2.9
通讯作者：
EKLUND, CM
中科院分区：
文献类型：
--
作者：
PRUSINER, SB;HADLOW, WJ;EKLUND, CM
根据小鼠脾组织的沉降谱，建立了一种从小鼠脾组织中部分纯化瘙痒病抗体的方法。差速离心法和去氧胆酸钠洗涤剂处理产生了一种命名为P5的组分，相对于细胞蛋白，该组分的瘙痒病传染性约为20倍。经EM检测，P5组分无细胞膜，但被核糖体严重污染。在蔗糖梯度中将组分P5离心至接近平衡时，羊瘙痒病的感染力分布在1.08-1.30克/厘米~3的密度范围内。平行率-区带分析表明，传染性分布在一个粒度范围内，S20，W[沉降系数为20度]。水中的C]来自.apprx的值。40 S至>；500S.在37或80度孵化P5。C，在破坏核糖体的条件下，极大地改变了试剂的速率-带状梯度分布。在这些条件下，该试剂以颗粒的形式与S20，W>；500 S一起沉淀。刮痕试剂在大小和密度方面的明显不均一性及其从一种形式转变为另一种形式的能力表明，该试剂的表面可能含有疏水结构域。
A procedure for the partial purification of the scrapie agent from mouse spleen was developed based on its sedimentation profile. Differential centrifugation and detergent treatment with sodium deoxycholate yielded a fraction designated P5 which was enriched for scrapie infectivity .apprx.20-fold with respect to cellular protein. The P5 fraction was devoid of cellular membranes but heavily contaminated with ribosomes as judged by EM. On centrifugation of the fraction P5 to near equilibrium in a sucrose gradient scrapie infectivity was distributed over a range of densities from 1.08-1.30 g/cm3. Parallel rate-zonal analysis showed that the infectivity was distributed over a range of particle sizes with S20,W [sedimentation coefficient at 20.degree. C in water] values from .apprx. 40 S to > 500 S. Incubation of P5 at 37 or 80.degree. C, under conditions that disrupt ribosomes, dramatically altered the rate-zonal gradient profile of the agent. Under these conditions, the agent sedimented as particles with S20,W > 500 S. The apparent heterogeneity of the scrapie agent with respect to both size and density and its ability to shift from 1 form to another suggest that the agent may contain hydrophobic domains on its surface.