A Chemical Modification- and Mass Spectrometry-Based Method for Thermodynamic Analysis of Protein-Ligand Binding

基于化学修饰和质谱的蛋白质-配体结合热力学分析方法

• 批准号: 0848462
• 负责人: Michael Fitzgerald
• 金额: $ 39万
• 依托单位: Duke University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0848462&HistoricalAwards=false
• 关键词: Chemical; Modification; Mass; Spectrometry; Based

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).The Divisions of Chemistry and Molecular and Cellular Biosciences support Professor Michael Fitzgerald of Duke University to develop and apply a mass spectrometry-based method known as SPROX (Stability of Proteins from Rates of Oxidation) to assess protein folding and the thermodynamic stability of protein-ligand complexes. Protocols are being optimized for model proteins and their ligands, and will then be extended to the analysis of protein-ligand binding in real biological mixtures (e.g., yeast cell lysates). The assay to be developed should have unmatched multiplex capabilities. It will create a unique opportunity to detect not only direct but also indirect binding, wherein binding of a target ligand to one protein either precludes or induces binding a second protein.The strategies and protocols to be created will be readily transferable to almost any protein-ligand system or biological mixture regardless of the ligand (i.e. small molecule, DNA, peptide, or other proteins) or binding affinity. The research provides interdisciplinary research experiences for both graduate and undergraduate students, and as well as high school students from the North Carolina Project SEED program, which is an on-going science enrichment program designed to steer disadvantaged, underrepresented minority students from North Carolina high schools into chemistry and chemistry-related careers.

该奖项是根据2009年的《美国恢复和再投资法》（公法111-5）资助的。化学，分子和细胞生物科学的划分支持杜克大学的教授迈克尔·菲茨杰拉德（Michael Fitzgerald）的教授，以开发和应用基于质谱的方法，称为SPROX（蛋白质的稳定性）（蛋白质与氧化速率）评估蛋白质互补的蛋白质互换性，并评估蛋白质的依从性。 方案正在针对模型蛋白及其配体进行优化，然后将其扩展到对实际生物混合物（例如酵母细胞裂解物）中蛋白质配体结合的分析。 要开发的测定应具有无与伦比的多重功能。 它将创造一个独特的机会，不仅可以直接检测间接结合，其中靶配体与一种蛋白质的结合是排除或诱导第二蛋白结合的蛋白质。要创建的策略和方案将很容易转移到几乎任何蛋白质 - 培养基系统或任何生物混合物中，无论是小小的蛋白质或生物学混合物（即小蛋白质），或其他蛋白质，或其他蛋白质，或其他蛋白质，或其他蛋白质，或其他蛋白质，或其他。 这项研究为研究生和本科生提供了跨学科的研究经验，以及北卡罗来纳州项目种子计划的高中生，这是一项正在进行的科学丰富计划，旨在将来自北卡罗来纳州北卡罗来纳州的弱势群体不足的少数族裔学生转向化学和化学与化学相关的职业。