Structure Function Analysis of the Multi-specific Drug Transporter OCT1

多特异性药物转运蛋白OCT1的结构功能分析

基本信息

批准号：
8666005
负责人：
SANJAY K NIGAM
金额：
$ 40.2万
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：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8666005
关键词：
ATP-Binding Cassette Transporters Adult Affect Area Binding Biological Assay Biology CCL21 gene Carrier Proteins Cations Cells Chemical Structure Chemicals Child Collaborations Computer Analysis Computer Simulation Crystallization Crystallography Data Data Collection Detergents Drug Design Drug Interactions Drug Kinetics Drug Transport Drug toxicity Epigenetic Process Epithelium Family Future Genetic Genetic Polymorphism Goals Homology Modeling In Vitro Integral Membrane Protein Knowledge Laboratories Lead Libraries Ligands Lipid Bilayers Lipids Mediating Membrane Proteins Metabolic Metformin Methods Modeling Molecular Neonatal Neurotransmitters Oats Oocytes Organ Organ Culture Techniques Organic Anion Transporters Organic Cation Transporter Organic Cation Transporter 1 POU2F1 gene Pathway interactions Permeability Pharmaceutical Preparations Pharmacologic Substance Phase Production Protein Family Proteins Publications Publishing Reaction Research Design Research Personnel Resolution Resources Route Running Science Slice Structural Protein Structure Testing Tissues Toxic effect Toxin United States National Institutes of Health Validation Xenobiotics base chemical property design high throughput screening in vivo in vivo Model inhibitor/antagonist innovation insight interest member molecular dynamics notch protein pharmacophore pressure protein purification public health relevance screening skills small molecule solute structural biology supercomputer synchrotron radiation three-dimensional modeling uptake virtual

项目摘要

DESCRIPTION (provided by applicant): Among the most highly expressed drug transporters in neonatal organs are the organic cation transporters (Octs), which transport drugs, neurotransmitters, metabolites and toxins. Many drug-drug interactions and metabolic derangements due to drugs are thought to occur at the level of the transporter, and certain polymorphisms can lead to drug toxicity. Oct1/SLC22A1 is the prototypical member of the SLC22A family of transporters and along with certain ABC transporters (e.g., MDR), Oats and Oatps, is one of the most important xenobiotic (phase III drug transporter) transporters in the body. The PI's lab identified Organic Anion Transporter 1 (Oat1/NKT/Slc22a6), which, along with Oct1 originally helped define the SLC22 transporter family. We have studied the in vivo function of several SLC22 transporters and performed detailed in vitro analysis and in silico modeling of substrate-transporter interactions. Nevertheless, detailed molecular analyses of substrate-transporter interactions-which are necessary to take a rational approach to diminishing adverse affects of drugs transported by Oct1-are limited by the absence of precise structural information. Here we propose to combine our knowledge of SLC22 transporter biology (and skill with in vitro, ex vivo and in vivo assays) together with the expertise of top-notch investigators in x-ray crystallography, protein and substrate modeling to investigate the molecular mechanisms of the key substrate-transporter interactions involved in Oct1-mediated organic cation xenobiotic handling. Although in this proposal we focus on Oct1, the results will also provide important information relevant to other SLC22 drug transporters. To our knowledge, we are among the very few groups in this field that has a proven ability to combine such diverse areas of expertise, including computationally biology, structural biology and in vivo/in vitro wet-lab biology and is one of the most innovative aspects of the proposed studies. We will crystallize and determine the high-resolution x-ray structure of Oct1 (SA1) in a continuing collaboration with Dr. Geoffrey Chang (involving both his laboratory and TransportPDB, a component of the NIH Protein Structural Initiative of which he is a founding member) to gain key insights into the structural basis of substrate binding and transport. We will use our documented expertise in ligand-based and transporter-based computational approaches (SA2) to identify the molecular determinants of substrates and the Oct1 protein mediating their interactions. Pharmacophore-based virtual screening of chemical structure libraries will be done, and identified structures will be used for molecular dynamic analyses to prioritize compounds. Identified compounds will be obtained and tested in wet-lab studies using in vitro, ex vivo and in vivo approaches well established in the PI's group (SA3). This multifaceted structural- computational-wet-lab strategy will result in major advances in understanding adverse drug reactions in children as well as adults and help move the drug transporter field forward.

描述（由申请人提供）：新生儿器官中最高表达的药物转运蛋白是有机阳离子转运蛋白（OCT），它们运输药物，神经递质，代谢物和毒素。人们认为许多由于药物引起的药物 - 药物相互作用和代谢危险在转运蛋白的水平上发生，某些多态性可能导致药物毒性。 OCT1/SLC22A1是SLC22A转运蛋白家族的典型成员，以及某些ABC转运蛋白（例如MDR），燕麦和燕麦是体内最重要的异种生物（III期药物转运蛋白）的燕麦群。 PI的实验室确定了有机阴离子转运蛋白1（OAT1/NKT/SLC22A6），该转运蛋白1（OAT1/NKT/SLC22A6）最初有助于定义SLC22转运蛋白家族。我们已经研究了几个SLC22转运蛋白的体内功能，并进行了详细的体外分析以及底物转运蛋白相互作用的计算机建模。然而，底物转运蛋白相互作用的详细分子分析是必须采用合理的方法来减少因缺乏精确的结构信息而受到限制的药物的不良影响。 Here we propose to combine our knowledge of SLC22 transporter biology (and skill with in vitro, ex vivo and in vivo assays) together with the expertise of top-notch investigators in x-ray crystallography, protein and substrate modeling to investigate the molecular mechanisms of the key substrate-transporter interactions involved in Oct1-mediated organic cation xenobiotic handling.尽管在此提案中，我们关注OCT1，但结果还将提供与其他SLC22药物转运蛋白有关的重要信息。据我们所知，我们是该领域中少数几个群体之一，具有合理的能力，可以将如此多样化的专业知识结合起来，包括计算生物学，结构生物学和体内/体外湿lab生物学，并且是拟议研究中最具创新性的方面之一。我们将在与Geoffrey Chang博士的持续合作中结晶并确定Oct1（SA1）的高分辨率X射线结构（涉及其实验室和TransportPDB，这是NIH蛋白结构倡议的组成部分，他是他是创始成员的组成部分），以使其成为基础结合和运输结构基础的关键见解。我们将在基于配体和基于转运蛋白的计算方法（SA2）中使用已记录的专业知识来识别底物的分子决定因素以及介导其相互作用的OCT1蛋白。将对化学结构文库进行基于药效团的虚拟筛选，并将确定的结构用于分子动态分析，以优先考虑化合物。使用体外，体内和体内方法在PI组中建立的（SA3）中，将在湿LAB研究中获得鉴定的化合物。这种多方面的结构计算-WET-LAB策略将在理解儿童和成年人的不良药物反应方面取得重大进展，并有助于将药物转运蛋白领域向前发展。