中文摘要：

最必要、关键的步在能递送的海绵状的 encephalopathy 的致病期间是细胞的 prion 蛋白质(PrP (C)) 的 conformational 变化到病理学的 isoform (PrP (Sc )) 。很多数据表明在房间表面的象 caveolae 一样领域(CLD ) 是 PrP 蛋白质的变换发生了的可能的地方。Apolipoprotein E (ApoE ) 是被认为由通过绑定形成蛋白质建筑群到位于房间表面的 clathrin 涂的坑的受体在 Alzheimer 的疾病和另外的 neurodegenerative 疾病的开发起一个重要作用的 apolipoprotein。在这研究，编码人的 ApoE3 的一个 914-bp cDNA 序列从 neuroblastoma 房间线 SH-SY5Y 被放大。三人的 ApoE 异构体从 Escherichia 关口 i 被表示并且净化。ApoE 特定的抗血清被与净化的 ApoE3 使兔子免疫准备。GST/His 下拉试金，免疫降水和 ELISA 表明三全身的 ApoE 异构体与 recombinant 交往在 vitro 的全身的 PrP 蛋白质。相应于蛋白质绑定的区域在 ApoE (氨基酸 1-194 ) 和 PrP (氨基酸 23-90 ) 的 N 终端的 N 终端片断被印射。而且， recombinant PrP 显示了能力从肝纸巾与本国的 ApoE 形成建筑群。我们的数据提供了在 ApoE 和 PrP 之间的分子的相互作用的直接证据。它也为从 PrP (C) 在 conformational 变换的过程在细胞的膜的表面上估计 CLD 的意义到 PrP (Sc ) 并且探查进能递送的海绵状的 encephalopathy 的致病供应了科学线索。

英文摘要：

The most essential and crucial step during the pathogenesis of transmissible spongiform encephalopathy is the conformational change of cellular prion protein （PrP^C） to pathologic isoform （prp^Sc）. A lot of data revealed that caveolae-like domains （CLDs） in the cell surface were the probable place where the conversion of PrP proteins happened. Apolipoprotein E （ApoE） is an apolipoprotein which is considered to play an important role in the development of Alzheimer＇s disease and other neurodegenerative diseases by forming protein complex through binding to the receptor located in the clathrin-coated pits of the cell surface. In this study, a 914-bp cDNA sequence encoding human ApoE3 was amplified from neuroblastoma cell line SH-SY5Y. Three human ApoE isomers were expressed and purified from Escherichia coli. ApoE-specific antiserum was prepared by immunizing rabbits with the purified ApoE3. GST/His pull-down assay, immunoprecipitation and ELISA revealed that three full-length ApoE isomers interact with the recombinant full-length PrP protein in vitro. The regions corresponding to protein binding were mapped in the N-terminal segment of ApoE （amino acid 1-194） and the N-terminal of PrP （amino acid 23-90）. Moreover, the recombinant PrP showed the ability to form a complex with the native ApoE from liver tissues. Our data provided direct evidence of molecular interaction between ApoE and PrP. It also supplied scientific clues for assessing the significance of CLDs on the surface of cellular membrane in the process of conformational conversion from PrP^C to PrP^Sc and probing into the pathogenesis of transmissible spongiform encephalopathy.