Basic studies on the biosynthesis of pathogenic prion protein

致病性朊病毒蛋白生物合成的基础研究

基本信息

批准号：
12460130
负责人：
MAKINO Sou-ichi
金额：
$ 9.09万
依托单位：
Obihiro University of Agriculture and Veterinary Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12460130/
关键词：
prion scrapie PrP conformational transformation glycosaminoglycan synthetic peptide BSE 伝達性海綿状脳症種特異性

项目摘要

An abnormal isoform of prion protein (PrPSc) is a major component of the infectious agent "prion" and the biosynthesis of PrPSc play a central role in pathogenesis of prion diseases. Once prion enters the host, PrPSc binds to normal prion protein (PrPC) and converts PrPC into PrPSc. During the conformational transformation, PrPSc acts as a template or seed. In this study, we analyzed inter-molecular interaction between PrPC and PrPSc that is a key event in PrPSc generation.We found that PrPC bound to heterologous PrPSc but did not convert to PrPSc and that the conversion reaction could be interfered by heterologous PrPC. These finding indicate that the conversion of PrPC to PrPSc is controlled more by the conformational transformation step. Analysis using mutant PrPC revealed that three amino acids locating central part of PrP is major determinant of the conversion reaction. We also examined the micro-environment where the conversion reaction takes place and found that sulfated glycans … More , a constituent of glycosaminoglycan, facilitated the PrPSc generation. Glycosaminoglycan is a major component of extracellular matrix so that environment in the outer surface of cells may be involved in the interaction between PrPC and PrPSc. Furthermore we found PrP synthetic peptide, aa119-141, 167-179 and 200-223 inhibited the PrPSc formation. These peptides formed high beta-sheet aggregates and bound to PrPC, implying the PrPC binding domain on the PrPSc molecule. In this study we have succeeded to produce an extensive panel of monoclonal antibodies against PrP molecule. The mAbs in this panel could be divided into at least 11 groups based on their epitope. Seven of them recognized linear epitopes on PrPC and/or denatured PrP molecule, while 3 of them recognized discontinuous epitopes on PrP molecule. The remaining one group of mAb, mAb 6H10 could differentiate PrPSc from PrPC. This extensive panel of mAb will be invaluable for further analyses of interaction between PrPSc and PrPC. Less

朊病毒蛋白 (PrPSc) 的异常异构体是传染性病原体“朊病毒”的主要成分，一旦朊病毒进入宿主，PrPSc 的生物合成在朊病毒疾病的发病机制中发挥着核心作用。并将 PrPC 转化为 PrPSc 在构象转化过程中，PrPSc 充当模板或种子。在本研究中，我们分析了 PrPC 和 PrPSc 之间的分子间相互作用。 PrPSc是PrPSc产生的关键事件。我们发现PrPC与异源PrPSc结合但没有转化为PrPSc，并且转化反应可能受到异源PrPC的干扰。这些发现表明PrPC向PrPSc的转化更多地受控。使用突变体 PrPC 的分析表明，位于 PrP 中心部分的三个氨基酸是转化反应发生的主要决定因素。硫酸化聚糖是糖胺聚糖的组成部分，促进了 PrPSc 的生成，糖胺聚糖是细胞外基质的主要成分，因此细胞外表面的环境可能参与 PrPC 和 PrPSc 之间的相互作用。 aa119-141、167-179 和 200-223 抑制这些肽的形成。 β-片层聚集并与 PrPC 结合，这意味着 PrPSc 分子上有 PrPC 结合域。在这项研究中，我们成功地产生了一系列针对 PrP 分子的单克隆抗体。该组中的 mAb 可以分为至少 11 组。其中 7 个识别 PrPC 和/或变性 PrP 分子上的线性表位，而其中 3 个识别 PrP 分子上的不连续表位。 mAb 组，mAb 6H10 可以区分 PrPSc 和 PrPC。这个广泛的 mAb 组对于进一步分析 PrPSc 和 PrPC Less 之间的相互作用非常有价值。