Structure and function of polyamine transporters

多胺转运蛋白的结构和功能

基本信息

批准号：
10615890
负责人：
Kenneth PK Lee
金额：
$ 42.95万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10615890
关键词：
Abnormal Cell Address Amyotrophic Lateral Sclerosis Autoimmune Diseases Autophagocytosis Binding Binding Sites Biochemical Biophysics C-terminal Cardiovascular Diseases Cations Cell Proliferation Cell membrane Cell physiology Cells Chemicals Clinical Complex Cryoelectron Microscopy Cysteine Cytosol Disease Drug Targeting Family Future Genetic Goals Health Hereditary Spastic Paraplegia Homeostasis Human Ion Channel Ion Pumps Knowledge Lipid Bilayers Lipid Binding Lipids Lumen of the Lysosome Lysosomes Maps Mediating Membrane Transport Proteins Metabolism Methods Molecular Molecular Conformation Mutagenesis Mutation Neurodegenerative Disorders Neuronal Ceroid-Lipofuscinosis Neurons PARK9 gene Parkinson Disease Pathway interactions Phosphorylation Physiology Play Polyamines Protein Dephosphorylation Proton Pump Pump Regulation Research Project Grants Resolution Role Route Signal Transduction Spermidine Spermine Stress Structure Syndrome System Testing Therapeutic Translations Transmembrane Transport Transport Process Vacuum Visualization Work cancer cell cytotoxic early onset human imaging improved inorganic phosphate insight loss of function mutation nanodisk neuron loss new therapeutic target novel therapeutic intervention operation particle pathogen phosphatidylinositol 3,5-diphosphate polycation prevent proteoliposomes rational design reconstitution theories therapeutic target three dimensional structure uptake

项目摘要

PROJECT SUMMARY Polyamines are a class of small organic polycations indispensable for many basic molecular and cellular processes including translation, electrical signaling, cell proliferation, and autophagy. Infectious and hyperproliferative diseases as well as many autoimmune, cardiovascular and neurodegenerative disorders are deeply connected to perturbations in polyamine abundance. Polyamine transport plays a major role in cellular polyamine homeostasis in both healthy and abnormal cells. Understanding the molecular basis of polyamine uptake and secretion has enormous potential to improve human health. However, despite decades of work, this subject continues to mystify. A critical barrier to deeper knowledge in polyamine transport is the complete absence of atomic structures of any polyamine transporter. The goal of this project is to elucidate the fundamental principles underlying polyamine transport and its regulation using a combination of structural and functional approaches. ATP13A2 is a lysosomal P-type ATP-driven pump tasked with the import of spermine and spermidine from the lysosome lumen to the cytosol. Mutations that cripple ATP13A2 function causes a spectrum of neurodegenerative diseases including Kufor-Rakeb syndrome, early-onset Parkinson’s disease, hereditary spastic paraplegia, neuronal ceroid lipofuscinosis and amyotrophic lateral sclerosis. ATP13A2 is, thus, a potential drug target. We have made significant inroads in our preliminary studies to determine high-resolution three-dimensional structures of human ATP13A2. In combination with functional analysis, these structures revealed ATP13A2’s luminal gating and polyamine selectivity mechanisms. Building on these preliminary results, we will leverage complementary electron cryo-microscopy, biophysical, biochemical, analytical chemical and mutagenesis strategies to further subject ATP13A2 to detailed mechanistic scrutiny. Specifically, we aim to investigate: 1) how lipids regulate ATP13A2 activity; 2) whether and how ATP13A2 pumps other cations into the lysosome; and 3) how ATP13A2 shuttles polyamines through the lipid bilayer. By addressing these questions, this research project will provide new insights into the basic operating and regulatory mechanisms of ATP13A2, which will broadly advance our understanding of polyamine transport and lysosome physiology. Structural and mechanistic discoveries from the proposed work may inform future rational design of therapeutics targeting ATP13A2.

项目概要多胺是一类小有机聚阳离子，对于许多基本分子和细胞来说是不可缺少的过程包括翻译、电信号传导、细胞增殖和自噬。过度增殖性疾病以及许多自身免疫性疾病、心血管疾病和神经退行性疾病与多胺丰度的扰动密切相关多胺运输在细胞中发挥着重要作用。了解健康和异常细胞中多胺稳态的分子基础。摄取和分泌在改善人类健康方面具有巨大潜力。然而，尽管经过数十年的努力，这一点仍然存在。深入了解多胺转运的一个关键障碍是完整的。不存在任何多胺转运蛋白的原子结构。该项目的目标是阐明其基本结构。基本原理多胺运输及其使用结构和功能相结合的调节 ATP13A2 是一种溶酶体 P 型 ATP 驱动泵，负责输入精胺和亚精胺从溶酶体腔到细胞质的突变会削弱 ATP13A2 的功能，从而产生谱图。神经退行性疾病，包括 Kufor-Rakeb 综合征、早发性帕金森病、遗传性因此，痉挛性截瘫、神经元蜡质脂褐质沉着症和肌萎缩侧索硬化症是一种疾病。我们在确定高分辨率的初步研究中取得了重大进展。结合功能分析，获得了人 ATP13A2 的三维结构。基于这些初步结果，揭示了 ATP13A2 的管腔门控和多胺选择性机制。我们将利用互补电子冷冻显微镜、生物物理、生物化学、分析化学和具体来说，我们的目标是进一步对 ATP13A2 进行详细的机制审查。研究：1) 脂质如何调节 ATP13A2 活性；2) ATP13A2 是否以及如何将其他阳离子泵入溶酶体；以及 3) ATP13A2 如何通过脂质双层运输多胺。该研究项目将为 ATP13A2 的基本运行和调节机制提供新的见解，这将广泛增进我们对多胺转运和溶酶体生理学的理解。拟议工作的机制发现可能会为未来治疗靶向的合理设计提供信息 ATP13A2。