Structure and function of polyamine transporters

多胺转运蛋白的结构和功能

• 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
• 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

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.

项目摘要 多胺是许多基本分子和细胞必不可少的小有机多阳离子 过程包括翻译，电信号传导，细胞增殖和自噬。传染性和 高增殖剂疾病以及许多自身免疫性，心血管和神经退行性疾病是 多胺转运在细胞中起主要作用 健康和异常细胞中的多胺稳态。了解多胺的分子基础 摄取和分泌具有改善人类健康的巨大潜力。但是，多斯特数十年来的工作，这 主题继续神秘。多胺传输中更深层次知识的关键障碍是完整的 没有任何多胺转运蛋白的原子结构。该项目的目的是阐明基本 使用结构和功能的组合，多胺传输的原理及其调节 方法。 ATP13A2是一种溶酶体P型ATP驱动的泵，其责任是精子的进口和 精子从溶酶体管腔到细胞质。削弱ATP13A2功能的突变会导致频谱 神经退行性疾病，包括Kufor-Rakeb综合征，早期帕金森氏病，遗传性 痉挛性截瘫，神经胶质脂肪肌动症和肌萎缩性侧索硬化症。因此，ATP13A2是 潜在的药物靶标。我们在初步研究中大大涉及确定高分辨率 人ATP13A2的三维结构。结合功能分析，这些结构 揭示了ATP13A2的腔门控和多胺选择性机制。以这些初步结果为基础 我们将利用完整的电子冷冻微镜，生物物理，生化，分析化学化学和 诱变策略将进一步对ATP13A2进行详细的机械审查。具体来说，我们的目标是 调查：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； 3）ATP13A2如何穿过脂质双层。通过解决这些问题， 该研究项目将为ATP13A2的基本操作和监管机制提供新的见解， 这将广泛提高我们对多胺传输和溶酶体生理的理解。结构和 提议的工作中的机械发现可能会为未来的治疗剂定位理性设计提供信息 ATP13A2。