Identification of an epitope in the C terminus of normal prion protein whose expression is modulated by binding events in the N terminus
Identification of an epitope in the C terminus of normal prion protein whose expression is modulated by binding events in the N terminus
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DOI：
10.1006/jmbi.2000.3986
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发表时间：
2000-08-18
影响因子：
5.6
通讯作者：
Sy, MS
中科院分区：
文献类型：
--
作者：
Li, RL;Liu, T;Sy, MS
我们的特点是一个面板的12个单克隆抗体（单克隆抗体）针对正常的人细胞朊蛋白（PrPC），使用ELISA和蛋白质印迹的重组PrP或合成肽片段的PrP的表位。第一组抗体由Mab 5 B2和8B 4代表，与PrP 23 -145反应，表明这些Mab的表位位于人PrP的23至145 N-末端区域。第二组包括Mab 1A 1、6 H3、7A 9、8 C6、8H 4、9 H7和2G 8。这些抗体结合定位于N-末端截短的重组PrP 90 -231内的表位。最后，Mab 5C 3、2C 9和7A 12识别PrP 23 -145和PrP 90 -231，表明该组的表位位于包含残基90至145的区域中。通过用PepSpot（TM）进行蛋白质印迹，所研究的单克隆抗体中只有三种（5 B2、8B 4和2G 8）与对应于人PrP片段的13个残基长的合成肽中存在的线性表位结合。其余9个单克隆抗体似乎识别构象表位。发现两种N末端特异性Mab阻止C末端特异性Mab 6 H3的结合。这一观察结果表明，非结构化的N-末端区域可能会影响朊病毒蛋白的折叠C-末端结构域内的局部构象。(C)北京大学出版社.
We have characterized the epitopes of a panel of 12 monoclonal antibodies (Mabs) directed to normal human cellular prion protein (PrPC) using ELISA and Western blotting of recombinant PrP or synthetic peptide fragments of PrP. The first group of antibodies, which is represented by Mabs 5B2 and 8B4, reacts with PrP23-145, indicating that the epitopes for these Mabs are located in the 23 to 145 N-terminal region of human PrP. The second group includes Mabs 1A1, 6H3, 7A9, 8C6, 8H4, 9H7 and 2G8. These antibodies bind to epitopes localized within N-terminally truncated recombinant PrP90-231. Finally, Mabs 5C3, 2C9 and 7A12 recognize both PrP23-145 and PrP90-231, suggesting that the epitopes for this group are located in the region encompassing residues 90 to 145. By Western blotting with PepSpot(TM) only three of Mabs studied (5B2, 8B4 and 2G8) bind to linear epitopes that are present in 13-residue long synthetic peptides corresponding to human PrP fragments. The remaining nine Mabs appear to recognize conformational epitopes. Two N terminus-specific Mabs were found to prevent the binding of the C terminus-specific Mab 6H3. This observation suggests that the unstructured N-terminal region may influence the local conformation within the folded C-terminal domain of prion protein. (C) 2000 Academic Press.