The regions of the Fe65 protein homologous to the phosphotyrosine interaction phosphotyrosine binding domain of Shc bind the intracellular domain of the Alzheimer's amyloid precursor protein
The regions of the Fe65 protein homologous to the phosphotyrosine interaction phosphotyrosine binding domain of Shc bind the intracellular domain of the Alzheimer's amyloid precursor protein
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DOI：
10.1074/jbc.270.52.30853
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发表时间：
1995-12-29
影响因子：
4.8
通讯作者：
Russo, T
中科院分区：
文献类型：
--
作者：
Fiore, F;Zambrano, N;Russo, T
Fe65是一种主要在哺乳动物神经系统的几个区域表达的蛋白质，蛋白质序列数据库搜索发现，Fe65含有两个磷酸酪氨酸相互作用(PID)或磷酸酪氨酸结合(PTB)结构域，这两个结构域以前在Shc接头分子中被发现。利用FE65的两个可能的PID/PTB结构域构建谷胱甘肽S-转移酶-FE65融合蛋白，共沉淀实验表明FE65的PID/PtB结构域与几个表观分子质量分别为135、115、105和51 kDa的蛋白相互作用。以FE65中含有PID/PTB结构域的区域为诱饵，用双杂交系统筛选人脑cDNA库，分离到三个不同的cDNA克隆，其中两个包含编码阿尔茨海默病β-淀粉样前体蛋白(APP)COOH端的cDNA片段，代表该膜蛋白的短胞内结构域，第三个克隆包含一个编码小鼠β-淀粉样前体蛋白的人类对应物COOH末端的cDNA片段。对筛选中发现的三个APP编码基因片段的比对表明，APP参与结合的区域集中在NPTY序列上，这与与Shc的PID/PTB结构域相互作用的生长因子受体的胞内域中的序列相似。
Fe65 is a protein mainly expressed in several districts of the mammalian nervous system, The search of protein sequence data banks revealed that Fe65 contains two phosphotyrosine interaction (PID) or phosphotyrosine binding (PTB) domains, previously identified in the Shc adaptor molecule. The two putative PID/PTB domains of Fe65 were used to construct glutathione S-transferase-Fe65 fusion proteins, Co-precipitation experiments demonstrated that the Fe65 PID/PTB domains interacted with several proteins of apparent molecular mass 135, 115, 105, and 51 kDa, The region of Fe65 containing the PID/PTB domains was used as a bait to screen a human brain cDNA library in yeast by the two-hybrid system, Three different cDNA clones were isolated, two of which contain overlapping segments of the cDNA encoding the COOH terminus of the Alzheimer's beta-amyloid-precursor protein (APP), that represents the short intracellular domain of this membrane protein, The third clone contains a cDNA fragment coding for the COOH terminus of the human counterpart of a mouse beta-amyloid-like precursor protein. The alignment of the three APP encoding cDNA fragments found in the screening suggests that the region of APP involved in the binding is centered on the NPTY sequence, which is analogous to that present in the intracellular domains of the growth factor receptors interacting with the PID/PTB domain of Shc.