STRUCTURAL CHARACTERIZATION OF PRION PROTEINS

朊病毒蛋白的结构表征

• 批准号: 6308820
• 负责人: STANLEY B PRUSINER
• 金额: $ 0.99万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6308820
• 关键词: Mammalia; aging; animal tissue; biological products; biomedical resource; congenital disorders; infection; nervous system; proteins; spectrometry; structural biology

Introduction: Prion diseases are unique fatal neurodgenerative diseases caused by novel pathogens termed prions. The major or only component of prions is an abnormal form of a normal cellular protein termed the prion protein (PrP). The normal form (PrPC) is converted to the pathogenic form (PrPSc) by an as yet unidentified posttranslational process. Mass spectrometry is essential for the detailed analysis of small-scale samples of both forms of the prion protein isolated from the brains of hamsters. Methods: Several novel methods have been refined for the analysis of PrPSc using LSIMS, ESIMS, MALDI and MS/MS for the characterization of peptides and oligosaccharides from enzyme digests. These analyses are complicated by the hydrophobic, insoluble and strongly aggregating propoerties, particularly of PrPSc. PrPC analyses have been delayed by problems of purifying the protein from brain. Preliminary results by MALDI show that the two forms are similar, but microheterogeneity has prevented any definitive conclusions being drawn. Comparisons are also required between PrPSc from different prion strains. Strain behavior could be due to differences in the N-linked oligosaccharides; perhaps cell specific lectins cause targetting of certain PrP species to a limited range of cell types, resulting in regional distribution of PrPSc. We plan to use LC/MS for much of this characterization. Discussion: Results obtained so far suggest that the modification of PrPC involved in the formation of PrPSc may be conformational rather than chemical. It is essential to confirm that PrpCand PrPSc either are or are not identical in order to develop hypotheses for the molecular mechanisms of prion diseases.

简介：王室疾病是独特的致命神经疾病 由新的病原体引起的疾病称为王室。 专业或唯一 Prions的成分是正常细胞蛋白的异常形式 称为Prion蛋白（PRP）。 正常形式（PRPC）转换 通过尚未确定的致病形式（PRPSC） 翻译后过程。 质谱对于 详细分析两种形式的小prion样本 从仓鼠的大脑中分离出来的蛋白质。 方法：几本小说 已经对使用LSIMS分析PRPSC的方法已被完善， ESIM，MALDI和MS/MS，用于表征肽和 酶消化的寡糖。 这些分析很复杂 通过疏水，不溶性和强烈聚集的建议， 特别是PRPSC。 PRPC分析已被问题延迟 从大脑中纯化蛋白质。 马尔迪的初步结果显示 这两种形式是相似的，但是微观均质性已经阻止了 得出的任何确定结论。 还需要比较 来自不同prion菌株的PRPSC之间。 压力行为可能是 由于N连接的寡糖差异；也许是细胞 特定的凝集素导致某些PRP物种靶向有限 细胞类型范围，导致PRPSC的区域分布。 我们 计划使用LC/MS进行大部分特征。 讨论： 到目前为止获得的结果表明，PRPC的修改涉及 在形成PRPSC的情况下可能是构象的，而不是化学的。 必须确认prpcand prpsc是或不是 相同的是为了发展分子机制的假设 病毒疾病。