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

描述（由申请人提供） 海洋污染通过我们接触来自海洋的受污染食物而对人类健康造成关切。人们仍然不太了解的是，为什么有些化学品具有持久性，在海洋生物体内积累，然后在人类体内积累，而其他化学品则没有。多药耐药（MDR）转运蛋白属于ATP结合盒（ABC）家族，是细胞内化学物质积累的主要生物学决定因素。 虽然它们被认为是环境化学品持久性的决定因素，并被用作预测药物供应和功效的工具，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，海胆和人类之间的异源物质清除转运蛋白的主要亚家族类型（ABCB，ABCC和ABCG）的功能显着保守。本申请探讨了这种保守的底物选择性的分子基础，作为第一步，转运蛋白生物学应用于预测污染物的持久性。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，开发其与主要海洋污染物相互作用的测定方法，并试图确定其高分辨率结构。他们将使用各向异性、ATP酶和全细胞分析来测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的保守残基和结构特征

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