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：
8707456
负责人：
GEOFFREY A CHANG
金额：
$ 17.24万
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/8707456
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 biophysical properties 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

描述（由申请人提供）海洋污染通过我们暴露于海洋中受污染的食物而关注的人类健康。仍然很少了解的是为什么有些化学物质持续存在，在海洋生物中积累，然后在人类中积累，而另一些则不是。属于ATP结合盒（ABC）家族的多药电阻（MDR）转运蛋白是细胞内化学积累的主要生物学决定因素。尽管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，异种生物的主要亚家族类型（ABCB，ABC和ABCG）的惊人功能保护消除了海胆和人之间的转运蛋白。该应用探讨了这种保守的底物选择性的分子基础，作为将转运蛋白生物学应用于污染物持久性预测的第一步。在这个项目中，调查人员将过度表达，溶解和净化海胆多饮出外排转运蛋白，开发与主要海洋污染物相互作用的测定法，并试图确定其高分辨率结构。他们将使用各向异性，ATPase和全细胞测定法测量与持续的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TransperPDB管道可用的哺乳动物转运蛋白的结构和功能进行比较，它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 Transporters多特异性结合袋中的底物相互作用至关重要