Detection of scrapie-associated fibril (SAF) proteins using anti-SAF antibody in non-purified tissue preparations.
Detection of scrapie-associated fibril (SAF) proteins using anti-SAF antibody in non-purified tissue preparations.
复制标题
使用非纯化组织制剂中的抗 SAF 抗体检测痒病相关原纤维 (SAF) 蛋白。
DOI：
10.1099/0022-1317-67-4-671
复制
发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Wisniewski,HM
中科院分区：
文献类型：
--
作者：
Rubenstein,R;Kascsak,RJ;Merz,PA;Papini,MC;Carp,RI;Robakis,NK;Wisniewski,HM
用抗皮肤瘙痒病相关纤维蛋白(SAF)的抗血清检测了3种不同的未纯化脑组织标本：突触体-线粒体部分、20%脑匀浆和用Sarkosyl提取的20%脑匀浆中的瘙痒病特异性多肽。负染电子显微镜检测到SAF蛋白在制剂中的浓度大于SAF的数量。这表明并不是所有的蛋白质都以SAF的形式存在。在正常和瘙痒病脑标本中都检测到免疫活性的33K到35K蛋白。在正常的脑组织标本中，该蛋白对完全的蛋白水解酶K(PK)具有易感性，因此推测，瘙痒病的感染是其翻译后修饰的原因，而翻译后修饰使其对PK产生抗性。这些修饰也可能在各种瘙痒病病原体的抗原性差异中发挥作用。在受瘙痒病影响的动物的脊髓和脾中也检测到SAF特异性蛋白。洗涤剂提取、PK处理和Western印迹分析是检测SAF蛋白的一种高度特异和敏感的方法。该方法可应用于病因不明的人类神经系统疾病。
Antisera raised to scrapie-associated fibril (SAF) proteins were used to detect scrapie-specific polypeptides in three different non-purified brain preparations: a synaptosomal-mitochondrial fraction, 20% brain homogenate and 20% brain homogenate extracted with Sarkosyl. The concentration of SAF proteins in the preparations was greater than the quantity of SAF as detected by negative stain electron microscopy. This suggests that not all of the protein exists in the form of SAF. An immunologically reactive 33K to 35K protein was detected in both normal and scrapie brain preparations. This protein was susceptible to complete proteinase K (PK) digestion in normal brain preparations and it is suggested that scrapie infection is responsible for post-translational modifications which confer PK resistance in scrapie preparations. These modifications may also play a role in the antigenic differences seen in a variety of scrapie agents. SAF-specific proteins were also detected in the spinal cords and spleens from scrapie-affected animals. Detergent extraction of material followed by PK treatment and Western blot analysis is a highly specific and sensitive method for the detection of SAF proteins. This procedure could be applied to human neurological diseases of unknown aetiology.