Studies of SLC41A1, a Mg2+-Transporting Protein Linked to Parkinson's Disease

SLC41A1（一种与帕金森病相关的 Mg2 转运蛋白）的研究

• 批准号: 10711860
• 负责人: Lejla Zubcevic
• 金额: $ 38.42万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711860
• 关键词: Artificial Membranes; Binding; Binding Sites; Biological Assay; Biophysics; Carrier Proteins; Cell Line; Cell physiology; Cells; Cellular Assay; Chemicals; Collaborations; Coupled; Cryoelectron Microscopy; Data; Disease; Ensure; Functional disorder; Genes; Grant; Health; Health system; Homeostasis; Housekeeping; Human; Investigation; Ion Channel; Ion Transport; Ions; Knock-out; Lipid Bilayers; Liposomes; Luciferases; Magnesium; Measures; Mediating; Membrane; Membrane Potentials; Metabolism; Mitochondria; Modeling; Molecular; Mutation; Nerve Degeneration; Neurobiology; Neuroblastoma; Neurodegenerative Disorders; Neurons; Oxidative Phosphorylation; Parkinson Disease; Pathogenesis; Permeability; Prevention strategy; Production; Proteins; Research; Resolution; Risk; Role; Sequence Alignment; Site-Directed Mutagenesis; Structure; Testing; Time; Water; Work; conformational conversion; cost; design; dopaminergic neuron; experimental study; fluorescence imaging; link protein; member; mitochondrial membrane; negative affect; new therapeutic target; novel; professor; reconstitution; solute; virtual

Project Summary/Abstract Magnesium has been largely overlooked in neurobiology as a “housekeeping” ion, but this view is directly challenged by evidence which shows that mutations in Mg2+ transporting proteins are associated with neurodegeneration. Mutations in the Mg2+ transporting protein SLC41A1 are associated with both increased and decreased risk of Parkinson's disease (PD). SLC41A1 might therefore be a causative gene for PD. Despite this, little is known about SLC41A1. For example, we do not yet know if it causes rapid changes in free cytosolic Mg2+ (like an ion channel) or if it works to maintain the levels of free cytosolic Mg2+ within the range required for normal cellular function (like a transporter). This distinction is required both to define the role of SLC41A1 in the Mg2+ homeostasis and its role in PD pathogenesis. Furthermore, a cellular role for SLC41A1 has not been determined and it is therefore not known how its dysfunction might lead to PD. We hypothesize that SLC41A1 is a Mg2+-permeable ion channel which, through its role in maintaining the cellular Mg2+ homeostasis, helps maintain the mitochondrial membrane potential and ensures that ATP production via oxidative phosphorylation proceeds undisturbed. In this application for a Stephen I. Katz ESI grant, we propose to use liposomal flux assays and lipid bilayer studies as well as electron cryo-microscopy (cryo-EM) to determine how SLC41A1 mediates Mg2+ transport, and to generate a model to determine its cellular role. Neuronal Mg2+ transport has thus far been virtually untouched by biophysical and structural investigation and the study proposed here represents the first steps towards understanding the molecular mechanisms of neuronal Mg2+ homeostasis as well as the Mg2+-dependent molecular origins of neurodegenerative disorders. In the long term, this study of SLC41A1 has the potential to lead to novel treatments of and preventative strategies against PD, a debilitating disease which costs the US health system more than $14 billion each year.

项目概要/摘要 镁作为一种“看家”离子在神经生物学中很大程度上被忽视了，但这种观点直接 受到证据的挑战，该证据表明 Mg2+ 转运蛋白的突变与 神经变性。 Mg2+ 转运蛋白 SLC41A1 的突变与 Mg2+ 的增加和 降低患帕金森病（PD）的风险。因此，SLC41A1 可能是 PD 的致病基因。尽管如此， 关于 SLC41A1 我们知之甚少。例如，我们还不知道它是否会导致游离胞质 Mg2+ 的快速变化 （如离子通道）或者它是否可以将游离胞质 Mg2+ 水平维持在正常所需的范围内 细胞功能（如转运蛋白）。需要这种区别来定义 SLC41A1 在 Mg2+ 中的作用 稳态及其在 PD 发病机制中的作用。此外，SLC41A1 的细胞作用尚未确定 因此尚不清楚其功能障碍如何导致帕金森病。 我们假设 SLC41A1 是一种 Mg2+ 渗透性离子通道，通过其维持 细胞 Mg2+ 稳态，有助于维持线粒体膜电位并确保 ATP 通过氧化磷酸化的生产不受干扰地进行。在本申请中，Stephen I. Katz ESI 授予，我们建议使用脂质体通量测定和脂质双层研究以及电子冷冻显微镜 （冷冻电镜）以确定 SLC41A1 如何介导 Mg2+ 转运，并生成模型以确定其细胞 角色。 神经元 Mg2+ 运输迄今为止几乎没有受到生物物理和结构研究的影响 这里提出的研究代表了理解分子机制的第一步 神经元 Mg2+ 稳态以及神经退行性疾病的 Mg2+ 依赖性分子起源。在 从长远来看，这项 SLC41A1 的研究有可能带来新的治疗和预防策略 PD 是一种使人衰弱的疾病，每年给美国卫生系统造成超过 140 亿美元的损失。