Structure and function of ABC transporters to understand persistence of global ma

描述(由申请人提供) 海洋污染是对人类健康的关切，因为我们接触到来自海洋的受污染的食物。人们仍然不太清楚的是，为什么一些化学物质是持久存在的，在海洋生物体内积累，然后在人类体内积累，而其他化学物质则不是。多药耐药(MDR)转运体属于三磷酸腺苷结合盒(ABC)家族，是细胞内化学积聚的主要生物学决定因素。虽然它们被认为是环境化学持久性的决定因素，并被用作预测药物可用性和有效性的工具，但它们尚未被系统地应用于预测污染物的持久性。研究人员的初步数据表明，海胆和人类之间的主要亚家族类型(ABCB、ABCC和ABCG)具有显著的功能保守，消除了海胆和人类之间的转运蛋白。这项应用探索了这种保守底物选择性的分子基础，作为应用转运蛋白生物学预测污染物持久性的第一步。在这个项目中，研究人员将过度表达、溶解和纯化 海胆多药外排转运蛋白，开发它们与主要海洋污染物相互作用的分析方法，并试图确定它们的高分辨率结构。他们将使用各向异性、ATPase和全细胞分析来衡量它们与持久性海洋污染物的相互作用。通过将海胆的结构和功能与已经通过TransportPDB管道获得的哺乳动物转运蛋白的结构和功能进行比较，他们将识别保守的残基和结构特征，这些残基和结构特征对于预测这些ABC转运蛋白的多特异性结合口袋中的底物相互作用至关重要

DESCRIPTION (provided by applicant) Marine pollution is of concern for human health through our exposure to contaminated food from the sea. What remains poorly understood is why some chemicals are persistent, accumulating in marine organisms and then in humans, while others are not. Multidrug resistance (MDR) transporters, belonging to the ATP Binding Cassette (ABC) family, are major biological determinants of intracellular chemical accumulation. While they have been implicated as determinants of environmental chemical persistence and used as tools for predicting availability and efficacy of drugs, they have yet to be systematically applied to predicting persistence of pollutants. The investigators' preliminary data indicate striking functional conservation of the major sub-family types (ABCB, ABCC and ABCG) of xenobiotic eliminating transporters between sea urchins and man. This application explores the molecular basis for this conserved substrate selectivity as a first step towards application of transporter biology to prediction of pollutant persistence. In this project, the investigators will over-express, solubilize, and purify sea urchin multidrug efflux transporter proteins, develop assays for their interaction with major marine pollutants and attempt to determine their high-resolution structures. They will measure their interaction with persistent marine pollutants using anisotropy, ATPase and whole cell assays. By comparing structure and functions of sea urchin with those of mammalian transporter proteins, already available through the TransportPDB pipeline, they will identify conserved residues and structural features that are essential for predicting substrate interaction in the poly-specific binding pocket of these ABC transporters