A MASS SPECTROMETER FOR PROTEIN FOOTPRINTING

用于蛋白质足迹分析的质谱仪

• 批准号: 8637341
• 负责人: MICHAEL L GROSS
• 金额: $ 44.99万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8637341
• 关键词: Address; Affinity; Alzheimer' s Disease; Amino Acids; Area; Binding; Brain Diseases; Calorimetry; Cereals; Chemicals; Commit; Communities; Complex; Data; Deuterium; Development; Fluorescence; Health; High Pressure Liquid Chromatography; Human; Hydrogen; Kinetics; Ligands; Malignant Neoplasms; Mass Spectrum Analysis; Measurement; Measures; Medicine; Membrane; Modification; Pain management; Peptides; Performance; Pharmaceutical Preparations; Photosynthesis; Play; Property; Protein Biochemistry; Protein Footprinting; Proteins; Proteomics; Research Support; Resolution; Resources; Science; Structure; System; Thermodynamics; Time; Universities; Virus Diseases; Washington; X-Ray Crystallography; base; improved; instrument; liquid chromatography mass spectrometry; mass spectrometer; oxidation; protein complex; protein folding; research study; technique development; tool

DESCRIPTION (provided by applicant): Mass spectrometry is a powerful tool for the identification of proteins and the determination of their structures and properties. The scientific community routinely employs MS-based proteomics. A new opportunity, protein footprinting, is emerging that takes advantage of these refined proteomics capabilities that combine high performance liquid chromatography and mass spectrometry. The development of these techniques is driven by a growing number of collaborative projects in biomedical science and other fields. The Washington University Mass Spectrometry Resource (WUMSR) has played a key role in the development of protein footprinting for more than a decade and is an ideal place to support its expansion. While it does not yet have the resolution of X-ray crystallography and NMR, MS-based footprinting is significantly more specific than the low resolution tools of CD, fluorescence, Raman, absorbance, SPR, and calorimetry. In addition footprinting occurs while the proteins and their complexes are in their native states, affording relevant measurements of many important thermodynamic and kinetic parameters. Footprinting approaches include hydrogen deuterium exchange (H/DX), fast photochemical oxidation of proteins (FPOP), and other amino acid specific chemical modifications. To fulfill and disseminate the promise of protein footprinting, we propose to add a high performance, LC/MS system and commit it full time to support this research. The proposed instrument will be used to address the needs of 17 collaborators who have problems in protein biochemistry and medicine. The proteins have implications in cancer, viral infections, pain management, and brain disorders, principally Alzheimer's disease (AD). There are implications as well as in energy and photosynthesis. The approaches will enable studies of protein folding and unfolding, permit the determination of interfaces between important peptides (e.g., A? with implications in AD), determine binding interfaces of small ligands including potential drugs, and measure the affinities of binding. We will extend these measurements to protein assemblies, including those that are membrane-bound. Given that the footprinting experiments produce data that "constrains" protein and protein-complex structure, just as in NMR, we predict that we can ultimately use these data to obtain "coarse-grained" structures, even in the absence of high resolution structural data. Progress in this area will impact a broad range of protein science with consequent effects on improving human health.

描述（由申请人提供）：质谱法是鉴定蛋白质并确定其结构和性质的有力工具。科学的 社区通常采用基于 MS 的蛋白质组学。蛋白质足迹分析这一新机会正在出现，它利用了这些结合了高效液相色谱和质谱的精细蛋白质组学功能。这些技术的发展是由生物医学科学和其他领域越来越多的合作项目推动的。十多年来，华盛顿大学质谱资源 (WUMSR) 在蛋白质足迹的发展中发挥了关键作用，是支持其扩展的理想场所。虽然尚不具备 X 射线晶体学和 NMR 的分辨率，但基于 MS 的足迹比 CD、荧光、拉曼、吸光度、SPR 和量热法等低分辨率工具具有更高的特异性。此外，当蛋白质及其复合物处于天然状态时，足迹就会发生，从而提供许多重要的热力学和动力学参数的相关测量。足迹方法包括氢氘交换 (H/DX)、蛋白质快速光化学氧化 (FPOP) 和其他氨基酸特异性化学修饰。为了实现和传播蛋白质足迹的承诺，我们建议添加高性能 LC/MS 系统，并全力支持这项研究。拟议的仪器将用于解决 17 位在蛋白质生物化学和医学方面存在问题的合作者的需求。这些蛋白质对癌症、病毒感染、疼痛管理和脑部疾病（主要是阿尔茨海默病（AD））具有重要意义。这对于能量和光合作用也有影响。这些方法将能够研究蛋白质折叠和解折叠，允许确定重要肽之间的界面（例如，与AD有关的Aβ），确定包括潜在药物在内的小配体的结合界面，并测量结合的亲和力。我们将把这些测量扩展到蛋白质组装体，包括那些膜结合的蛋白质组装体。鉴于足迹实验产生的数据“约束”蛋白质和蛋白质复合物结构，就像在 NMR 中一样，我们预测，即使在没有高分辨率结构数据的情况下，我们最终也可以使用这些数据来获得“粗粒度”结构。该领域的进展将影响广泛的蛋白质科学，从而改善人类健康。