Bioanalytical techniques and affinity assays for DNA damage and molecular interactions

这项研究计划将专注于开发分析技术和超灵敏的分析方法，用于分析微量的DNA损伤，并研究涉及G蛋白偶联受体(GPCRs)的相互作用。各种内源性和环境性因素可损伤人类DNA，导致各种不利的健康后果。因此，检测DNA损伤是至关重要的。目前使用抗体的免疫分析方法不能检测多种类型的DNA损伤。这项拟议的研究将使用DNA修复蛋白来识别和结合多种类型的DNA损伤。结合毛细管电泳分离和激光诱导荧光检测，新的亲和分析将能够检测与环境、职业和临床暴露相关的广泛DNA损伤，如紫外线辐射、癌症治疗药物和其他环境致癌物。 GPCRs调节我们对激素、神经递质和环境刺激物的大部分生理反应。人们对GPCRs的结构进行了大量的研究，但对GPCRs的动态相互作用知之甚少。这项研究将建立独特的激光诱导荧光偏振分析毛细管电泳法来研究GPCRs与其配体相互作用的构象变化和结合结构域。这些分析和研究对治疗发展是有用的，因为GPCRs代表了被证明具有治疗价值的最大类别的分子靶点。 这项研究将加强我们在生物分析化学方面的卓越成就，促进跨学科合作，并培训多学科人员来应对生化和环境研究面临的分析挑战。

This research program will focus on developing analytical techniques and ultrasensitive assays for analysis of minute amounts of DNA damage and for studying interactions involving G-protein-coupled receptors (GPCRs). A variety of endogenous and environmental agents can damage human DNA, leading to various adverse health outcomes. Thus, it is crucial to detect DNA damage. Current immunoassays using antibodies cannot detect multiple types of DNA damage. The proposed research will use DNA repair proteins to recognize and bind to multiple types of DNA damage. Combined with capillary electrophoresis separation and laser induced fluorescence detection, the new affinity assays will enable detection of a wide range of DNA damage that is relevant to environmental, occupational, and clinical exposure, such as UV radiation, cancer treatment drugs, and other environmental carcinogens. GPCRs mediate most of our physiological responses to hormones, neurotransmitters and environmental stimulants. Much effort has been directed to determining structures of GPCRs, but little is understood about the dynamic interactions The proposed research will establish unique laser induced fluorescence polarization assays with capillary electrophoresis separation to study the conformation changes and binding domains of GPCRs interactions with their ligands. These assays and studies are useful to therapeutic development because GPCRs represent the largest class of molecular targets with proven therapeutic value. This research will strengthen our established excellence in bioanalytical chemistry, foster inter-disciplinary collaborations, and train multidisciplinary people to address analytical challenges facing biochemical and environmental studies.