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Structural and functional studies of the TRPM2 channel

TRPM2通道的结构和功能研究

基本信息

批准号：
10604261
负责人：
Juan Du
金额：
$ 41.56万
依托单位：
VAN ANDEL RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604261
关键词：
ADP Receptors Adenosine Diphosphate Ribose Affect Agonist Allosteric Site Alzheimer&apos s Disease Apoptosis Architecture Binding Binding Sites Biological Biological Assay Biophysics Bipolar Disorder Blood Vessels Body Temperature Brain Brain Injuries C-terminal Calcium Cations Cell Death Cell membrane Cell physiology Cells Characteristics Coiled-Coil Domain Complex Consensus Cryoelectron Microscopy Curcumin Cyclic ADP-Ribose Data Development Diabetes Mellitus Disease Edetic Acid Electrophysiology (science)Excision Family Family member Fever Foundations Heart Heating Homologous Gene Human Hypothalamic structure Immune Immune response Inflammation Insulin Ion Channel Ion Channel Protein Ions Knowledge Length Ligand Binding Ligands Link Lysosomes Maps Membrane Metabolic Metabolism Mission Mitochondria Molecular Molecular Conformation Molecular Structure Mutation N-terminal Nerve Neurodegenerative Disorders Nucleotides Oxidative Stress Pathologic Permeability Pharmaceutical Preparations Pharmacology Physiologic Thermoregulation Physiological Physiological Processes Play Positioning Attribute Process Property Protons Public Health Research Research Project Grants Resolution Rest Rhizome Role Sampling Site Smooth Muscle Solid Sorting Specificity Stroke Structure Therapeutic Agents Time Tissues Tumeric United States National Institutes of Health Work X-Ray Crystallography Zebrafish antagonist density experimental study improved insight insulin secretion interdisciplinary approach novel novel therapeutics particle prevent process improvement pyrophosphatase receptor sensor simulation small molecule sulfated glycoprotein 2

项目摘要

Body temperature is strictly maintained in a narrow range to protect the delicate nerves in the brain and other body tissues, because improper body temperature gives rise to fever, brain injury, and stroke. TRPM2 is the major warmth-sensing receptor in the brain regulating core body temperature and preventing overheating as fever occurs. TRPM2 is a Ca2+-permeable, nonselective ion channel that is highly expressed in brain but is also found in the heart, vascular and smooth muscle, and immune cells. It is uniquely activated by Ca2+ and ADP ribose (ADPR), a product of the metabolism of NAD+ and a secondary messenger released upon oxidative stress. The activation of TRPM2 results in both Ca2+ entry across the plasma membrane and Ca2+ release from lysosomes. Therefore, TRPM2 plays fundamental role in Ca2+-dependent array of physiological processes and cellular functions from insulin secretion to immune response to cell death. It has been implicated in Alzheimer disease, stroke, and other neurodegenerative diseases. TRPM2 belongs to the TRPM (melastatin-like transient receptor potential) subfamily of the TRP superfamily. Despite sharing the characteristic TRPM N-terminal homology regions (MHRs) and C-terminal coiled-coil domains, TRPM2 is uniquely assembled with a C-terminal NHDT9-H domain, a homolog to the human mitochondrial ADP-ribose pyrophosphatase NUDT9. Functional studies, including binding assays, electrophysiology, and molecular simulations, provided a consensus view that ADPR binds to the NUDT9-H domain, but proof of the ADPR binding site is lacking, and the molecular basis for the action of the agonist ADPR on TRPM2 in the presence of calcium remains unknown. The gating of TRPM2 is further modulated by many molecules and ions that range from protons to nucleotides (cyclic ADPR, AMP, 8-Br-cADPR) to curcumin (which is isolated from rhizomes of Curcuma longa), acting by way of multiple mechanisms. At present, we don't know where these molecules and ions bind to TRPM2 or how they activate the channel or modulate its function. We have obtained two cryo-EM structures of zebrafish TRPM2 in the apo/closed and ADPR/Ca2+-bound open state, with the latter representing the first active state of TRPM family members. We identified a novel ADPR binding site that is located outside the NUDT9-H domain and was completely unknown before. Building on this preliminary data, we propose to continue the structural studies of TRPM2 combined with complementary electrophysiology experiments, binding assays, and X-ray crystallography, which will define the molecular basis for a comprehensive gating mechanism and pharmacology. These advances will provide a solid foundation for developing new drugs against neurodegenerative diseases and for a deeper understanding the function of the entire TRPM family.

严格地将体温保持在一个狭窄的范围内，以保护大脑和其他部位脆弱的神经身体组织，因为不适当的体温会引起发烧、脑损伤和中风。TRPM2是大脑中主要的温觉感受器，调节核心体温，防止过热就会发烧。TRPM2是一种钙离子通透性的非选择性离子通道，在大脑中高度表达，但在心脏、血管和平滑肌以及免疫细胞中也有发现。它唯一地被钙离子激活，并且腺苷二磷酸核糖(ADPR)，是NAD+代谢的产物，是一种在氧化应激。TRPM2的激活导致钙离子跨质膜进入和钙离子内流从溶酶体中释放。因此，TRPM2在钙离子依赖的生理信号通路中起着基础性作用。从胰岛素分泌到对细胞死亡的免疫反应的过程和细胞功能。一直以来与阿尔茨海默病、中风和其他神经退行性疾病有关。 TRPM2属于色氨酸蛋白超家族中的TRPM(Melastatin样瞬时受体电位)亚家族。尽管共享特征的TRPM N-末端同源区域(MHR)和C-末端螺旋线圈结构域，TRPM2唯一地与C-末端NHDT9-H结构域组装，该结构域与人类的同源物线粒体ADP-核糖焦磷酸酶NUDT9。功能研究，包括结合分析，电生理学和分子模拟提供了ADPR与NUDT9-H结合的一致观点结构域，但缺乏ADPR结合位点的证据，以及激动剂作用的分子基础钙离子存在时TRPM2上的ADPR尚不清楚。对TRPM2的选通进行进一步调制由许多分子和离子组成，从质子到核苷酸(环ADPR、AMP、8-溴-cADPR)到姜黄素(从姜黄根状茎中分离出来)，通过多种机制发挥作用。在… 目前，我们不知道这些分子和离子与TRPM2结合的位置，也不知道它们如何激活通道或调节其功能。我们获得了斑马鱼TRPM2在apo/闭合和ADPR/Ca~(2+)结合的开放状态下的两种冷冻-EM结构，后者代表了TRPM家族成员的第一活性状态。我们发现了一个新的ADPR结合位点，该结合位点位于NUDT9-H结构域外，以前完全未知。在这些初步数据的基础上，我们建议继续进行TRPM2的结构研究，结合互补的电生理学实验、结合分析和X射线结晶学，这将为全面的门控机制和药理学确定分子基础。这些进展将为我们提供坚实的为开发治疗神经退行性疾病的新药和加深了解奠定基础整个TRPM系列的功能。