Wille H, Drewes G, Biernat J, Mandelkow E M, Mandelkow E
Max-Planck-Unit for Structural Molecular Biology, Hamburg, Germany.
J Cell Biol. 1992 Aug;118(3):573-84. doi: 10.1083/jcb.118.3.573.
Recent evidence from several laboratories shows that the paired helical filaments of Alzheimer's disease brains consist mainly of the protein tau in an abnormally phosphorylated form, but the mode of assembly is not understood. Here we use EM to study several constructs derived from human brain tau and expressed in Escherichia coli. All constructs or tau isoforms are rodlike molecules with a high tendency to dimerize in an antiparallel fashion, as shown by antibody labeling and chemical crosslinking. The length of the rods is largely determined by the region of internal repeats that is also responsible for microtubule binding. One unit length of the repeat domain (three or four repeats) is around 22-25 nm, comparable to the cross-section of Alzheimer PHF cores. Constructs corresponding roughly to the repeat region of tau can form synthetic paired helical filaments resembling those from Alzheimer brain tissue. A similar self-assembly occurs with the chemically cross-linked dimers. In both cases there is no need for phosphorylation of the protein.
来自多个实验室的最新证据表明,阿尔茨海默病大脑中的双螺旋丝主要由异常磷酸化形式的tau蛋白组成,但组装方式尚不清楚。在这里,我们使用电子显微镜研究了几种源自人类大脑tau蛋白并在大肠杆菌中表达的构建体。所有构建体或tau异构体都是棒状分子,通过抗体标记和化学交联表明,它们具有以反平行方式二聚化的高度倾向。棒的长度很大程度上由内部重复区域决定,该区域也负责与微管结合。重复结构域的一个单位长度(三个或四个重复)约为22 - 25纳米,与阿尔茨海默病双螺旋丝核心的横截面相当。大致对应于tau蛋白重复区域的构建体可以形成类似于阿尔茨海默病脑组织的合成双螺旋丝。化学交联的二聚体也会发生类似的自组装。在这两种情况下,蛋白质都无需磷酸化。