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

ABC转运蛋白的结构和功能，以了解全球MA的持久性

基本信息

批准号：
8550059
负责人：
GEOFFREY A CHANG
金额：
$ 17.43万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-24 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8550059
关键词：
ABCC1 gene ATP Hydrolysis ATP phosphohydrolase ATP-Binding Cassette Transporters Animals Anisotropy Antisense Oligonucleotides Binding Biological Biological Assay Biological Models Biology Carrier Proteins Cell Line Cell physiology Cells Chemicals Complementary DNA Crystallization Data Embryo Employee Strikes Environment Exposure to Family Fishes Fluorescence Fluorescence Anisotropy Fundulus heteroclitus Genetic Goals Health Human In Vitro Integral Membrane Protein Laboratories Marines Measures Mediating Membrane Proteins Modeling Molecular Molecular Structure Multi-Drug Resistance Mus Mussels Mytilus Pattern Pharmaceutical Preparations Pollution Polychlorinated Biphenyls Proteins Recombinants Research Infrastructure Research Personnel Resolution Sea Sea Urchins Seafood Structure Substrate Interaction System Toxic effect United States National Institutes of Health Xenobiotics Yeasts Zebrafish aquatic organism base bioaccumulation cytotoxicity drug efficacy embryo cell environmental chemical experience expression cloning in vitro Assay in vitro testing in vivo inhibitor/antagonist loss of function man marine organism molecular transporter multi drug transporter organochlorine pesticide pollutant pollutant interaction protein structure public health relevance tool uptake

项目摘要

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 Public Health Relevance: Certain molecular transporters found in sea urchins and other marine organisms determine the uptake of persistent pollutants from the sea. The goal of this high-impact project is to provide a detailed molecular framework for understanding the structure and function of these important transporters. This application fulfills an important need to develop biologically-based tools to predict bioaccumulation and toxicity of marine pollutants as relevant to human health and environment.

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影响人类健康。人们仍然知之甚少的是，为什么有些化学物质具有持久性，在海洋生物体内积累，然后在人类体内积累，而另一些则不然。多药耐药性 (MDR) 转运蛋白属于 ATP 结合盒 (ABC) 家族，是细胞内化学积累的主要生物决定因素。虽然它们被认为是环境化学持久性的决定因素，并用作预测药物的可用性和功效的工具，但它们尚未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，海胆和人类之间的外源消除转运蛋白的主要亚科类型（ABCB、ABCC和ABCG）具有显着的功能保守性。该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，开发其与主要海洋污染物相互作用的测定方法，并尝试确定其高分辨率结构。他们将使用各向异性、ATP 酶和全细胞测定来测量它们与持久性海洋污染物的相互作用。通过比较海胆与已通过 TransportPDB 管道获得的哺乳动物转运蛋白的结构和功能，他们将识别保守残基和结构特征，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公共卫生相关性：海胆和其他海洋生物中发现的某些分子转运蛋白决定了对海洋中持久性污染物的吸收。这个具有高影响力的项目的目标是提供详细的分子框架，以了解这些重要转运蛋白的结构和功能。该应用满足了开发基于生物学的工具来预测与人类健康和环境相关的海洋污染物的生物累积和毒性的重要需求。