The Role of ZIP12 in Zinc Homeostasis and Associated Neurodegenerative Pathologies

ZIP12 在锌稳态和相关神经退行性病理学中的作用

基本信息

  • 批准号:
    10559709
  • 负责人:
  • 金额:
    $ 18.89万
  • 依托单位:
  • 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:
    2022-02-01 至 2025-01-31
  • 项目状态:
    未结题

项目摘要

Zinc is an essential micro-nutrient that participates in catalytic and structural functions touching nearly every metabolic process in the cell. While alterations in neuronal Zn2+ distribution have been associated with multiple disease states including Alzheimer’s Disease (AD), ALS, schizophrenia, and depression, the molecular mechanism of Zn2+ homeostasis in neurons is largely unexplored. The Zinc and Iron-reg- ulated transport Proteins (ZIP) mediate the entrance of first row transition metals into the cytoplasm. hZIP12, one of the fourteen human ZIPs, is a Zn2+ uptake transporter expressed in the brain. hZIP12 cSNPs (coding single nucleotide polymorphisms) have been associated with AD and abnormalities in the caudate nucleus. Importantly, our previous structural functional studies of the homologous hZIP4 enables postulation of the likely functional alteration introduced by observed cSNP. This proposal is based on the hypothesis that studies of hZIP12 and variants associated with neurodegenerative diseases will serve as an entrance point to build testable mechanistic models of neuronal Zn2+ homeostasis. Aim one tests the hypothesis that hZIP12 mutations associated with neuronal diseases impact transport activity and/or sur- face expression. hZIP12-mediated Zn2+ transport will be measured in neuronal SH-SY5Y cells expressing wild type and mutated proteins. Membrane targeting and dimerization studies will characterize predicted structural effects of observed cSNPs. The structural and functional consequence of three types of vari- ants will be examined: 1) one cSNP linked to AD, 2) two cSNPs associated with morphological alterations of the caudate nucleus; and 3) two cSNPs hypothesized to alter metal selectivity. Aim two tests the hy- pothesis that hZIP12 dysfunction modifies cellular Zn2+ pools and metal distribution. Cytosolic Zn2+ levels, as well as transmembrane transporter expression, will be assessed under early steady state and non- toxic Zn2+ levels. To this end, targeted as well as unbiased global approaches will be employed: 1) size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS) profiling of the proteome will be combined with mass spectrometry (LC-MS/MS) to identify Zn2+ binding proteins. 2) The expression of compensatory genes participating in the response will be evaluated via unbiased approaches (RNA-Seq) as well as directed Q-PCR of genes known to be involved in Zn2+ homeostasis/distribution. Results from these studies will initiate the definition of molecular and subcellular mechanisms of Zn2+ homeostasis in the brain, provide a detailed description of hZIP12 function in neuronal Zn2+ subcellular distribution, and elucidate the role of hZIP12 in relevant neurodegenerative diseases.
锌是一种必需的微量营养素,参与催化和结构功能,几乎触及 细胞中的每一个代谢过程。虽然神经元Zn 2+分布的改变与 患有多种疾病状态,包括阿尔茨海默病(AD)、ALS、精神分裂症和抑郁症, 神经元中Zn 2+内稳态的分子机制在很大程度上未被探索。锌和铁的关系 稳定转运蛋白质(ZIP)介导第一排过渡金属进入细胞质。 hZIP 12是人类14种ZIP之一,是在脑中表达的Zn 2+摄取转运蛋白。hZIP12 cSNPs(编码单核苷酸多态性)与AD和阿尔茨海默病相关。 尾状核重要的是,我们以前对同源hZIP 4的结构功能研究使得 假设由观察到的cSNP引入的可能的功能改变。这项建议是根据 假设hZIP 12和与神经退行性疾病相关的变体的研究将作为 一个入口点,以建立可测试的机制模型的神经元锌2+稳态。目标一测试 假设与神经元疾病相关的hZIP 12突变影响转运活性和/或表面活性, 面部表情hZIP 12介导的Zn 2+转运将在表达以下蛋白的神经元SH-SY 5 Y细胞中测量: 野生型和突变蛋白质。膜靶向和二聚化研究将表征预测的 观察到的cSNPs的结构效应。三种类型的瓦里的结构和功能后果- 将检查蚂蚁:1)一个与AD相关的cSNP,2)两个与形态学改变相关的cSNP 尾状核;和3)两个cSNP假设改变金属选择性。目的二是检验hy- 假设hZIP 12功能障碍改变细胞Zn 2+池和金属分布。胞浆Zn 2+水平, 以及跨膜转运蛋白表达,将在早期稳态和非稳态下进行评估。 锌离子中毒为此,将采用有针对性和无偏见的全球方法: 排阻色谱-电感耦合等离子体质谱(SEC-ICP-MS)分析 蛋白质组将与质谱法(LC-MS/MS)结合以鉴定Zn 2+结合蛋白。2)的 参与应答的补偿基因的表达将通过无偏的 方法(RNA-Seq)以及已知参与Zn 2+的基因的定向Q-PCR 稳态/分布。这些研究的结果将启动分子和 锌2+稳态在脑中的亚细胞机制,提供了hZIP 12的详细描述 在神经元Zn 2+亚细胞分布中的作用,并阐明hZIP 12在相关的 神经退行性疾病

项目成果

期刊论文数量(3)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Heterologous Expression of Full-Length and Truncated Human ZIP4 Zinc Transporter in Saccharomyces cerevisiae.
  • DOI:
    10.3390/biom12050726
  • 发表时间:
    2022-05-21
  • 期刊:
  • 影响因子:
    5.5
  • 作者:
  • 通讯作者:
The transmembrane domains mediate oligomerization of the human ZIP4 transporter in vivo.
  • DOI:
    10.1038/s41598-022-24782-6
  • 发表时间:
    2022-12-06
  • 期刊:
  • 影响因子:
    4.6
  • 作者:
    Liu Y;Bafaro EM;Cowan AE;Dempski RE
  • 通讯作者:
    Dempski RE
Single-molecule quantification of the oligomeric state of ZIP transporters in mammalian cells with fluorescence correlation spectroscopy.
利用荧光相关光谱法对哺乳动物细胞中 ZIP 转运蛋白寡聚状态进行单分子定量。
  • DOI:
    10.1016/bs.mie.2023.04.021
  • 发表时间:
    2023
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Liu,Yuting;Bafaro,ElizabethM;Dempski,RobertE
  • 通讯作者:
    Dempski,RobertE
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Robert Edward Dempski其他文献

Robert Edward Dempski的其他文献

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{{ truncateString('Robert Edward Dempski', 18)}}的其他基金

The role of ZIP12 in zinc homeostasis and associated neurodegenerative pathologies
ZIP12 在锌稳态和相关神经退行性疾病中的作用
  • 批准号:
    10452802
  • 财政年份:
    2022
  • 资助金额:
    $ 18.89万
  • 项目类别:
The molecular determinants of zinc uptake mediated by hZIP4
hZIP4介导的锌摄取的分子决定因素
  • 批准号:
    9432517
  • 财政年份:
    2014
  • 资助金额:
    $ 18.89万
  • 项目类别:
The molecular determinants of zinc uptake mediated by hZIP4
hZIP4介导的锌摄取的分子决定因素
  • 批准号:
    8631438
  • 财政年份:
    2014
  • 资助金额:
    $ 18.89万
  • 项目类别:
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