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部门支持杜克大学的Michael Fitzgerald教授开发和应用一种基于质谱的方法，称为SPROX（氧化率的蛋白质稳定性），以评估蛋白质折叠和蛋白质-配体复合物的热力学稳定性。 实验方案正在针对模型蛋白及其配体进行优化，然后将扩展到分析真实的生物混合物中的蛋白-配体结合（例如，酵母细胞裂解物）。 待开发的检测试剂盒应具有无与伦比的多重检测能力。 这将创造一个独特的机会，不仅检测直接，而且间接的结合，其中结合的靶配体的一种蛋白质，要么排除或诱导结合的第二个蛋白质。的策略和协议将容易转移到几乎任何蛋白质-配体系统或生物混合物，无论配体（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提供了跨学科的研究经验，以及高中学生从北卡罗来纳州项目种子计划，这是一个正在进行的科学富集计划，旨在引导弱势，代表性不足的少数民族学生从北卡罗来纳州高中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.