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

该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。化学和分子与细胞生物科学系支持杜克大学的Michael Fitzgerald教授开发和应用一种基于质谱的方法，称为SPROX（蛋白质从氧化速率的稳定性），以评估蛋白质折叠和蛋白质配体复合物的热力学稳定性。目前正在对模型蛋白质及其配体的方案进行优化，然后将扩展到实际生物混合物（例如酵母细胞裂解物）中蛋白质-配体结合的分析。待开发的检测方法应具有无与伦比的多重功能。它将创造一个独特的机会，不仅可以检测直接结合，还可以检测间接结合，其中靶配体与一种蛋白质的结合可以阻止或诱导与另一种蛋白质的结合。要创建的策略和方案将很容易地转移到几乎任何蛋白质-配体系统或生物混合物中，而不考虑配体（即小分子，DNA，肽或其他蛋白质）或结合亲和力。该研究为研究生和本科生以及来自北卡罗来纳项目SEED项目的高中生提供了跨学科的研究经验，该项目是一个正在进行的科学充实计划，旨在引导北卡罗来纳高中的弱势、代表性不足的少数民族学生进入化学和化学相关的职业。

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.