Structure and Function of Heme-based Dioxygenases

血红素双加氧酶的结构和功能

基本信息

批准号：
9973608
负责人：
Syun-Ru Yeh
金额：
$ 41.92万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9973608
关键词：
Address Binding Binding Sites Biochemical Biological Process Biophysics Clinical Clinical Treatment Clinical Trials Data Diet Dioxygenases Disease Drug Binding Site Drug Targeting Enzymes Essential Amino Acids Exhibits Family Funding Goals Group Structure Guidelines Heme Human Knowledge Kynurenine Malignant Neoplasms Mass Spectrum Analysis Mental Depression Molecular Neurodegenerative Disorders Outcome Study Pathway interactions Pharmaceutical Preparations Physiological Physiology Property Protein Isoforms Public Health Reaction Role Site Structure Structure-Activity Relationship Sulfamethoxazole Techniques Technology Testing Therapeutic Therapeutic Intervention Tryptophan Tryptophan 2,3 Dioxygenase Tumor Escape X-Ray Crystallography base biophysical techniques cancer cell cancer immunotherapy design disorder control disorder prevention drug development indoleamine inhibitor/antagonist innovation insight member new therapeutic target novel screening small molecule three dimensional structure

项目摘要

SUMMARY Tryptophan (Trp) is the least abundant essential amino acid. The majority of our dietary Trp is metabolized through the kynurenine (KYN) pathway. The first and rate-limiting step of the KYN pathway is catalyzed by three heme-based dioxygenases, tryptophan dioxygenase (hTDO), indoleamine 2,3-dioxygenase 1 (hIDO1), and indoleamine 2,3-dioxygenase 2 (hIDO2). Recently it was found that the three dioxygenases are expressed in cancer cells to promote cancer immune escape. Consequently they have been considered as key drug targets for cancer immunotherapy. Despite their importance, the structural and functional properties of these enzymes remain elusive, which has hindered the progress of the field. The central hypothesis of this project, as supported by our preliminary data, is (i) the functional properties of the three dioxygenases are regulated by cellular metabolites and (ii) each dioxygenase exhibits distinct structural features and possesses unique drug binding sites. We will test our hypothesis by addressing two specific aims: (i) identify cellular metabolites that interact with each dioxygenase and define the related regulatory mechanisms, and (ii) define structural differences between the three dioxygenases and determine new small molecule binding sites in each dioxygenase. We will use a new high- throughput mass spectrometry-based screening technology to identify metabolites that interact with each dioxygenase and use X-ray crystallography and spectroscopic techniques to define their specific molecular interactions and functional consequences. These studies will reveal previously unknown cellular players in dioxygenase-related human physiology that may impact the specific functions of these enzymes in cancer and other diseases, thereby offering novel information enabling innovative molecular approaches for disease prevention and control. In parallel, we will use an integrated approach, involving a wide spectrum of biochemical and biophysical techniques, and a group of structurally diverse inhibitors as probes to define unique structural features and new small molecule binding sites in each dioxygenase. The outcome of these studies will offer important knowledge enabling better understanding of structure-and-function relationships of the three heme- based dioxygenases and expanding our toolkit for rational design of enzyme-selective inhibitors. We have assembled a team of experts to carry out this innovative project with the multifaceted approach. These studies will address significant gaps in our knowledge of molecular mechanisms underlying the biological functions of the three dioxygenases and provide important new insights into related drug development and disease treatment.

概括色氨酸 (Trp) 是含量最少的必需氨基酸。我们饮食中的色氨酸大部分被代谢通过犬尿氨酸 (KYN) 途径。 KYN 途径的第一步也是限速步骤是由三种基于血红素的双加氧酶：色氨酸双加氧酶 (hTDO)、吲哚胺 2,3-双加氧酶 1 (hIDO1) 和吲哚胺 2,3-双加氧酶 2 (hIDO2)。最近发现这三种双加氧酶在促进癌细胞免疫逃逸。因此它们被认为是关键的药物靶点癌症免疫疗法。尽管它们很重要，但这些酶的结构和功能特性仍然存在难以捉摸，阻碍了该领域的进步。该项目的中心假设得到了我们的支持初步数据是（i）三种双加氧酶的功能特性受细胞代谢物调节 (ii) 每种双加氧酶表现出独特的结构特征并拥有独特的药物结合位点。我们将通过解决两个具体目标来检验我们的假设：（i）识别与每个目标相互作用的细胞代谢物双加氧酶并定义相关的调节机制，以及（ii）定义三者之间的结构差异双加氧酶并确定每种双加氧酶中新的小分子结合位点。我们将使用新的高基于通量质谱的筛选技术，用于识别与每种代谢物相互作用的代谢物双加氧酶并使用 X 射线晶体学和光谱技术来定义其特定分子相互作用和功能后果。这些研究将揭示以前未知的细胞参与者与双加氧酶相关的人体生理学可能会影响这些酶在癌症和癌症中的特定功能其他疾病，从而提供新的信息，从而实现疾病的创新分子方法预防和控制。与此同时，我们将采用综合方法，涉及广泛的生化和生物物理技术，以及一组结构多样的抑制剂作为探针来定义独特的结构每种双加氧酶的特征和新的小分子结合位点。这些研究的结果将提供重要的知识可以更好地理解三种血红素的结构和功能关系基于双加氧酶并扩展我们的工具包以合理设计酶选择性抑制剂。我们有组建了一个专家团队，以多方面的方式开展这一创新项目。这些研究将解决我们对生物功能背后的分子机制的认识上的重大差距三种双加氧酶，并为相关药物开发和疾病治疗提供重要的新见解。