STUDY OF THE PROTEIN IN MINK ENCEPHALOPATHY

水貂脑病中蛋白质的研究

• 批准号: 3122592
• 负责人: RICHARD FLOYD MARSH
• 金额: $ 9.91万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-29 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3122592
• 关键词: Alzheimer's disease; amyloid proteins; antibody; electron microscopy; endopeptidases; gel electrophoresis; hamsters; high performance liquid chromatography; histopathology; ion exchange chromatography; neoplastic cell culture for noncancer research; phospholipase C; posttranslational modifications; prions; protein purification; protein sequence; protein structure function; solubility; spongiform encephalopathy; tissue /cell culture

The long term objectives of this application are to determine how to prion protein becomes modified in infected animals, and how this modification may produce cell injury. Investigation of a new incident of transmissible mink encephalopathy (TME) has resulted in the identification of two distinct disease syndromes in hamsters. The first has an incubation period of sixty-five days and has the predominant clinical signs of hyperesthesia and cerebellar ataxia. The second syndrome has an incubation period of 160 days and is clinically characterized by lethargy with no hyperesthesia or cerebellar ataxia. Each syndrome has been "cloned" by serial passage at high dilution. Examination of the prion protein (PrP) in each of these syndromes has revealed differences in solubilities, protease sensitivities, and molecular weights. This is the first time that PrP differences have been seen in two distinct syndromes in a single outbred species. Therefore, the differences in PrP observed can not be due to changes in primary amino acid sequence, but rather to posttranslational modification. The specific aims of this proposal are to biochemically characterize the PrPs from the two syndromes in an attempt to identify conformational and/or other posttranslational modifications to these proteins which may effect their processing within the cell and influence the disease syndrome produced. Furthermore, it may be possible to determine if the different PrPs induce their associated syndromes in a "strain-specific" manner by testing other infected species (mink, cattle, squirrel monkeys) for specific PrP isoforms. This proposal is relevant to human dementias of the Alzheimer's type because it will attempt to determine biochemical differences in the amyloid-like prion protein from two distinct clinical syndromes in hamsters. Changes identified may explain differences observed in the solubility, protease sensitivity, and molecular weights of the proteins. These findings may further relate to changes in protein processing in the brain which influence the course and type of disease produced.

该应用程序的长期目标是确定如何朊病毒 受感染动物体内的蛋白质发生改变，以及这种改变是如何发生的 可能会产生细胞损伤。 一起新的传染性水貂脑病（TME）事件的调查 导致了两种不同疾病综合征的鉴定 仓鼠。 第一个病毒的潜伏期为六十五天， 主要临床症状为感觉过敏和小脑性共济失调。 这 第二综合征潜伏期为160天，临床上 特点是嗜睡，无感觉过敏或小脑性共济失调。 每个 综合征已通过高稀释度连续传代被“克隆”。 对每种综合征中的朊病毒蛋白 (PrP) 进行检查 揭示了溶解度、蛋白酶敏感性和分子水平的差异 重量。 这是首次在两种药物中发现 PrP 差异 单一近交物种的独特综合症。 因此，差异 观察到的 PrP 不可能是由于一级氨基酸序列的变化， 而是翻译后修饰。 该提案的具体目标是从生化角度表征 来自两种综合征的 PrP，试图识别构象和/或 这些蛋白质的其他翻译后修饰可能会影响 它们在细胞内的处理并影响疾病综合征 产生的。 此外，还可以确定不同的 PrPs 通过“菌株特异性”方式诱导其相关综合征 检测其他受感染物种（水貂、牛、松鼠猴） 特定的 PrP 亚型。 该提案与阿尔茨海默氏症类型的人类痴呆症相关 因为它将尝试确定生物化学差异 来自两种不同临床综合征的淀粉样蛋白样朊病毒蛋白 仓鼠。 确定的变化可以解释观察到的差异 溶解度、蛋白酶敏感性和蛋白质的分子量。 这些发现可能进一步与蛋白质加工的变化有关 影响疾病产生的过程和类型的大脑。