Singer David, Lehmann Jörg, Hanisch Katja, Härtig Wolfgang, Hoffmann Ralf
Bioanalytics, Center for Biotechnology and Biomedicine (BBZ), Faculty of Chemistry and Mineralogy, University of Leipzig, Deutscher Platz 5, 04103 Leipzig, Germany.
Biochem Biophys Res Commun. 2006 Aug 4;346(3):819-28. doi: 10.1016/j.bbrc.2006.05.201. Epub 2006 Jun 9.
Neurofibrillary tangles, which represent a major pathological hallmark in Alzheimer's disease (AD), are deposits of the hyperphosphorylated microtubule-associated tau protein (PHF-tau). However, a link between the phosphorylation pattern and the cause or the progress of AD is still missing. The work reported here focused on PHF-tau specific local phosphorylation patterns at Thr212/Ser214 and Thr231/Ser235 using monoclonal antibodies (mAb) generated against correspondingly modified peptides. The binding motifs of the obtained six mAbs were characterized with non-, mono-, and double-phosphorylated peptides as well as terminally shortened sequences. Five mAbs stained neurofibrillary tangles, neuritic plaques, and neuropil threads from autoptic brains of AD cases. Four mAbs recognized PHF-tau without significant cross-reactivity towards normal human tau, bovine tau, and dephosphorylated PHF-tau in ELISA and Western blot analysis. Thus, double phosphorylation is sufficient to distinguish PHF-tau from all other tau versions and there is no need to postulate any PHF-tau specific conformation for this region.
神经原纤维缠结是阿尔茨海默病（AD）的主要病理标志，是高度磷酸化的微管相关tau蛋白（PHF-tau）的沉积物。然而，磷酸化模式与AD病因或进展之间的联系仍然缺失。本文报道的工作聚焦于使用针对相应修饰肽产生的单克隆抗体（mAb），研究Thr212/Ser214和Thr231/Ser235处PHF-tau特异性局部磷酸化模式。用非磷酸化、单磷酸化和双磷酸化肽以及末端缩短序列对获得的六种mAb的结合基序进行了表征。五种mAb对AD病例尸检脑的神经原纤维缠结、神经炎性斑块和神经毡丝进行了染色。在ELISA和蛋白质印迹分析中，四种mAb识别PHF-tau，对正常人tau、牛tau和去磷酸化的PHF-tau无明显交叉反应。因此，双磷酸化足以将PHF-tau与所有其他tau变体区分开来，无需假定该区域存在任何PHF-tau特异性构象。
