BIOCHEMICAL AND BIOPHYSICAL CHARACTERIZATION OF PRION PROTEIN 2D CRYSTALS

朊病毒蛋白二维晶体的生物化学和生物物理表征

• 批准号: 8169775
• 负责人: STANLEY B PRUSINER
• 金额: $ 0.35万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169775
• 关键词: Benign; Biochemical; Computer Retrieval of Information on Scientific Projects Database; Cryoelectron Microscopy; Disease; Dose; Excision; Funding; Grant; Institution; Mass Spectrum Analysis; Molecular Conformation; Neurodegenerative Disorders; PrPSc Proteins; Preparation; Prions; Protein Isoforms; Proteins; Protocols documentation; Research; Research Personnel; Resources; Sampling; Scrapie; Source; Staining method; Stains; Structure; United States National Institutes of Health; base; particle; polyacrylamide gels; protein purification

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Prions cause transmissible, neurodegenerative diseases with invariably fatal progression. A prion is an infective particle consisting exclusively of the prion protein. Two forms are discriminated: the benign, cellular form PrPC, and the disease-causing "scrapie" isoform PrPSc, which is confirmationally rearranged. Since PrPC and PrPSc differ in structure, studies of each conformation are essential. Our apporach focuses on PrPSc. We developed a high-yield protein purification protocol. Our preparations contain prion protein 2D crystals, which are structurally analyzed with standard and low-dose cryo electron microscopy. However, to further the quality and purity of our samples, we need to identify contaminants/copurified proteins from our samples for their systematic removal. This is the task for mass spectrometry. On a collaborative basis, copurified proteins are to be identified by analysis of bands derived from Coomassie-stained polyacrylamide gels.

这个子项目是众多研究子项目之一