Innovative Membrane Protein Tech: Biophysical Character

创新膜蛋白技术：生物物理特性

• 批准号: 6962724
• 负责人: PETER KUHN
• 金额: $ 15.59万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-24 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6962724
• 关键词: X ray crystallography; biomedical automation; biophysics; biotechnology; electron microscopy; functional /structural genomics; high throughput technology; membrane proteins; method development; nanotechnology; nuclear magnetic resonance spectroscopy; peptide chemical synthesis; structural biology; synthetic protein; technology /technique development

Aim 3. Adapt the available high throughput biophysical characterization techniques for IMPs, evaluate the quality of IMP production and sample preparation, and correlate data. a. Electron Microscopy - develop protein/lipid/detergent combinations by automated EM for sample homogeneity and protein quality by single particle picking using a low voltage EM (LVEM). b. Nuclear Magnetic Resonance Spectroscopy- develop the use of NMR probes and 1D 1H, 2D 1H15N and site specific 2H and l5N labeling with TROSY NMR for characterizing and optimizing IMP preparations. c. X-ray - adapt current automated nanovolume crystallization and imaging technologies for IMP use and evaluate IMP quality for potential crystallization and diffraction characterization.

目的3.将现有的高通量生物物理表征技术应用于IMP，评价IMP的生产和样品制备的质量，并对数据进行关联。 A.电子显微镜-通过使用低电压EM(LVEM)通过单颗粒拾取，通过自动化EM开发蛋白质/脂肪/洗涤剂组合，以确保样品的均一性和蛋白质质量。 B.核磁共振波谱-开发使用核磁共振探针和1D 1H、2D 1H15N以及TROSY核磁共振位点特定的2H和15N标记来表征和优化IMP制剂。 C.X射线-适应当前用于IMP的自动纳米体积结晶和成像技术，并评估IMP的质量，以进行潜在的结晶和衍射表征。