MOLECULAR MECHANISMS FOR PRION PROTEIN CONVERSION AND CELL-TYPES INVOLVED IN THE CONVERSION IN VIVO

朊病毒蛋白体内转化的分子机制和参与转化的细胞类型

• 批准号: 09470154
• 负责人: KATAMINE Shigeru
• 金额: $ 7.04万
• 依托单位: NAGASAKI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470154/
• 关键词: PRION DISEASES; PRION PROTEIN; PEPTIDES; THERAPEUTICS; PROTEIN-PROTEIN INTERACTION; ファージディスプレイ; ノックアウトマウス; 神経細胞死; 発症予防; ペプチドライブラリ; 遺伝子改変マウス; ニューロン; グリア; プリオン; 遺伝子改変動物; プルキンエ細胞; アストロサイト; 脱髄; 軸索

Post-translational conversion of prion protein (PrP) is thought to be the nature of prion propagation. In the present study, we identified aberrant mRNA species in the brain of an ataxic mouse line, Ngsk PrnpィイD10/0ィエD1, lacking PrP. These mRNAs are chimeric between the non-coding exons l/2 of PrP gene (Prnp) and the novel sequence encoding PrP-like protein (PrPLP), a putative membrane glycoprotein with 22% identity with PrP in the primary amino acid structure. Rabbit antiserum raised against recombinant PrPLP specifically immunoprecipitated a protein (m.w.: 19 kD) from lysates of A293T cells transfected with an expression vector. Physical association of PrPLP with PrP was demonstrated in both cell-free system (IAsys cuvette) and transfected cells, suggesting a functional interaction of the two proteins. In addition, by screening a random peptide (12-mer) library, we identified 3 synthetic peptides with a potential of high affinity binding to PrP. Interestingly, one of them reduced the level of the proteinase K-resistant pathogenic form of PrP in vitro with a dose-dependent manner. Both PrPLP and the PrP-binding peptides would be valuable reagents to elucidate molecular mechanisms for the PrP conversion and allow for the development of therapeutics of prion diseases.

prion蛋白（PRP）的翻译后转化被认为是pr传播的本质。在本研究中，我们发现了低调小鼠系的大脑中的异常mRNA物种Ngsk Prnpei D10/0E D1，缺乏PRP。这些mRNA是PRP基因（PRNP）L/2的非编码外显子与编码PRP样蛋白（PRPLP）的新序列之间的嵌合。用表达矢量翻译。 PRPLP与PRP的物理关联均在无细胞系统（IASYS比色杯）和翻译细胞中证明，这表明两种蛋白质的功能相互作用。此外，通过筛选随机肽（12-mer）文库，我们确定了3个合成胡椒体，具有高亲和力与PRP的潜力。有趣的是，其中一个以剂量依赖性方式降低了蛋白酶K抗抗性的致病形式。 PRPLP和PRP结合的胡椒体都将是阐明PRP转化率的分子机制并允许开发Prion疾病疗法的有价值的试剂。

项目成果

Nishida, N. et al.: "A mouse prion protein (PrP) transgene rescues Purkinje cell degeneration and demyelination in mice deficient for PrP"Lab. Invest.. 79. 689-697 (1999)

Nishida, N. 等人：“小鼠朊病毒蛋白 (PrP) 转基因可挽救 PrP 缺陷小鼠的浦肯野细胞变性和脱髓鞘”实验室。

Moore,R.C.et al.: "Ataxia in prion protein (PrP) deficient mice associated with upregulation of the novel PrP-like protein doppel"J.Mol.Biol.. 292(4). 797-817 (1999)

Moore, R.C. 等人：“朊病毒蛋白 (PrP) 缺陷小鼠中的共济失调与新型 PrP 样蛋白二元的上调相关”J.Mol.Biol.. 292(4)。

Katamine,S.Nishida,N. 他: "Impaired motor coordination in mice lacking prion protein." Cell.Mol.Neurobiol.18. 731-742 (1998)

Katamine, S. Nishida, N. 等人：“缺乏朊病毒蛋白的小鼠运动协调受损。”Cell.Mol.Neurobiol.18 (1998)。

Satoh,J.et al.: "Cultured skin fibroblasts isolated from mice devoid of the prion prote in gene express major heat shock proteins in response to heat stress"Exp.Neurol.. 151. 105-115 (1998)

Satoh, J.等人：“从基因中缺乏朊病毒蛋白的小鼠中分离出的培养皮肤成纤维细胞在响应热应激时表达主要热休克蛋白”Exp.Neurol.. 151. 105-115 (1998)

Moore, R. C. et al.: "Ataxia in prion protein (PrP) deficient mice associated with upregulation of the novel PrP-like protein doppel"J. Mol. Biol.. 292(4). 797-819 (1999)

Moore, R. C. 等人：“朊病毒蛋白 (PrP) 缺陷小鼠中的共济失调与新型 PrP 样蛋白二元体的上调相关”J.