A PREPARATION OF ALZHEIMER PAIRED HELICAL FILAMENTS THAT DISPLAYS DISTINCT TAU-PROTEINS BY POLYACRYLAMIDE-GEL ELECTROPHORESIS
A PREPARATION OF ALZHEIMER PAIRED HELICAL FILAMENTS THAT DISPLAYS DISTINCT TAU-PROTEINS BY POLYACRYLAMIDE-GEL ELECTROPHORESIS
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DOI：
10.1073/pnas.87.15.5827
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发表时间：
1990-08-01
影响因子：
11.1
通讯作者：
DAVIES, P
中科院分区：
文献类型：
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作者：
GREENBERG, SG;DAVIES, P
成对螺旋丝(PHF)是阿尔茨海默病(AD)神经原纤维缠结(NFT)的重要组成部分。我们没有分离NFT，而是选择了可以从AD大脑匀浆中提取的PHF种群。约50%的PHF免疫反应可在27,200次内获得。G在含有0.8M氯化钠的缓冲液中均质后的上清液。我们通过利用PHF在两性离子洗涤剂和2-巯基乙醇存在下的不溶性，通过0.45-微米过滤器过滤去除聚集体，以及蔗糖密度离心法，进一步丰富了PHF。富含PHF的组份含有两到五个57-68 kDa的蛋白质，表现出与PHF相同的抗原性。因为57到68 kDa的PHF蛋白在抗原性上与tau相关。蛋白质，它们类似于tau。以前在非傅立叶变换中观察到的蛋白质。然而，进一步的分析表明，与PHF相关的tau。可以与正常的、可溶的.tau区分开来。由不识别人类成人tau的PHF抗体产生。以及一维和二维的页面。
Paired helical filaments (PHFs) are prominent components of Alzheimer disease (AD) neurofibrillary tangles (NFTs). Rather than isolating NFTs, we selected for PHF populations that can be extracted from AD brain homogenates. About 50% of PHF immunoreactivity can be obtained in 27,200 .times. g supernatants following homogenization in buffers containing 0.8 M NaCl. We further enriched for PHFs by taking advantage of their insolubility in the presence of zwiterionic detergents and 2-mercaptoethanol, removal of aggregates by filtration through 0.45-.mu.m filters, and sucorse density centrifugation. PHF-enriched fractions contained two to five proteins of 57-68 kDa that displayed the same antigenic properties as PHFs. Since the 57- to 68-kDa PHF proteins are antigenically related to .tau. proteins, they are similar to the .tau. proteins previously observed in NFTs. However, further analysis revealed that PHF-associated .tau. can be distinguished from normal, soluble .tau. by PHF antibodies that do not recognize human adult .tau. and by one- and two-dimensional PAGE.