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

• 批准号: 9020978
• 负责人: SANJAY K NIGAM
• 金额: $ 38.84万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9020978
• 关键词: ATP-Binding Cassette Transporters; Adult; Affect; Area; Avena sativa; 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; Health; 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; 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 Genes; 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; novel therapeutics; pharmacophore; pressure; protein purification; screening; skills; small molecule; solute; structural biology; supercomputer; synchrotron radiation; three-dimensional modeling; uptake; virtual

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）：新生儿器官中表达最高的药物转运蛋白是有机阳离子转运蛋白（Octs），其转运药物、神经递质、代谢物和毒素。许多药物间相互作用和由药物引起的代谢紊乱被认为发生在转运蛋白水平上，并且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，与某些 ABC 转运蛋白（例如 MDR）、燕麦和 Oatps 一起，是体内最重要的外源性（III 期药物转运蛋白）转运蛋白之一。 PI 的实验室鉴定了有机阴离子转运蛋白 1 (Oat1/NKT/Slc22a6)，它与 ​​Oct1 最初一起帮助定义了 SLC22 转运蛋白家族。我们研究了几种 SLC22 转运蛋白的体内功能，并对底物-转运蛋白相互作用进行了详细的体外分析和计算机建模。然而，对底物-转运蛋白相互作用的详细分子分析——这对于采取合理的方法来减少 Oct1 转运药物的不利影响是必要的——由于缺乏精确的结构信息而受到限制。在这里，我们建议将我们的 SLC22 转运蛋白生物学知识（以及体外、离体和体内测定的技能）与 X 射线晶体学、蛋白质和底物建模方面的顶尖研究人员的专业知识相结合，以研究 SLC22 转运蛋白的分子机制。 Oct1介导的有机阳离子异生素处理中涉及的关键底物-转运蛋白相互作用。尽管在本提案中我们重点关注 Oct1，但结果也将提供与其他 SLC22 药物转运蛋白相关的重要信息。据我们所知，我们是该领域极少数能够结合如此不同专业领域（包括计算生物学、结构生物学和体内/体外湿实验室生物学）的团队之一，并且是最具创新性的团队之一拟议研究的各个方面。我们将与 Geoffrey Chang 博士（涉及他的实验室和 TransportPDB）持续合作，结晶并确定 Oct1 (SA1) 的高分辨率 X 射线结构，TransportPDB 是 NIH 蛋白质结构计划的组成部分，他是该计划的创始成员）以获得对底物结合和运输的结构基础的重要见解。我们将利用我们在基于配体和基于转运蛋白的计算方法 (SA2) 方面的专业知识来确定底物的分子决定因素以及介导它们相互作用的 Oct1 蛋白。将进行基于药效基团的化学结构库虚拟筛选，识别出的结构将用于分子动力学分析，以确定化合物的优先级。将使用 PI 小组 (SA3) 成熟的体外、离体和体内方法在湿实验室研究中获得和测试已鉴定的化合物。这种多方面的结构-计算-湿实验室策略将在了解儿童和成人药物不良反应方面取得重大进展，并有助于推动药物转运蛋白领域的发展。

项目成果

Kidney versus Liver Specification of SLC and ABC Drug Transporters, Tight Junction Molecules, and Biomarkers.

SLC 和 ABC 药物转运蛋白、紧密连接分子和生物标志物的肾脏与肝脏规格。

• 发表时间: 2016
• 作者: Martovetsky, Gleb;Bush, Kevin T;Nigam, Sanjay K
• 通讯作者: Nigam, Sanjay K

What do drug transporters really do?

药物转运蛋白的真正作用是什么？

• DOI: 10.1038/nrd4461
• 发表时间: 2015-01
• 期刊: Nature Reviews Drug Discovery
• 影响因子: 120.1
• 作者: Nigam SK

Handling of Drugs, Metabolites, and Uremic Toxins by Kidney Proximal Tubule Drug Transporters.

• DOI: 10.2215/cjn.02440314
• 发表时间: 2015-11-06
• 期刊: Clinical journal of the American Society of Nephrology : CJASN
• 作者: S. Nigam;Wei Wu;K. Bush;M. Hoenig;R. Blantz;V. Bhatnagar
• 通讯作者: V. Bhatnagar

The organic anion transporter (OAT) family: a systems biology perspective.

有机阴离子转运蛋白（OAT）家族：系统生物学视角。

• DOI: 10.1152/physrev.00025.2013
• 发表时间: 2024-09-13
• 期刊: Physiological reviews
• 影响因子: 33.6
• 作者: S. Nigam;K. Bush;Gleb Martovetsky;Sun;Henry C. Liu;E. Richard;V. Bhatnagar;Wei Wu
• 通讯作者: Wei Wu