Ksiezak-Reding H, Yen S H
Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461.
Neuron. 1991 May;6(5):717-28. doi: 10.1016/0896-6273(91)90169-z.
Highly purified and SDS-soluble paired helical filaments (PHFs) were immunogold labeled and immunoblotted with antibodies to tau: Tau 14 (N-terminal half), AH-1 (microtubule-binding domain), and Tau 46 (C-terminal end). The main component of PHFs was modified tau of 68, 64, and 60 kd, also called A68 or PHF-tau. Trypsin digestion reduced the maximum width of PHFs by 10%-20%, increased aggregation of filaments, and abolished the binding of Tau 14, but had no effect on the binding of AH-1. The smallest tau-reactive tryptic fragments were 13 and 7-8 kd, positive with AH-1, and negative with Tau 46. Our results and the model of Crowther and Wischik suggest that by self-association and anti-parallel arrangement of the microtubule-binding domains, PHF-tau forms the backbone of PHFs.
高度纯化且可溶于十二烷基硫酸钠(SDS)的双螺旋丝(PHFs)用抗tau蛋白的抗体进行免疫金标记和免疫印迹分析:Tau 14(N端半段)、AH - 1(微管结合结构域)和Tau 46(C端)。PHFs的主要成分是68、64和60千道尔顿的修饰tau蛋白,也称为A68或PHF - tau。胰蛋白酶消化使PHFs的最大宽度减少了10% - 20%,增加了丝状物的聚集,并消除了Tau 14的结合,但对AH - 1的结合没有影响。最小的与tau反应的胰蛋白酶片段为13以及7 - 8千道尔顿,与AH - 1呈阳性反应,与Tau 46呈阴性反应。我们的结果以及Crowther和Wischik的模型表明,通过微管结合结构域的自缔合和反平行排列,PHF - tau形成了PHFs的主干。