THE ROLE OF TMEM163 IN ZINC HOMEOSTASIS

TMEM163 在锌稳态中的作用

• 批准号: 10287961
• 负责人: MATH P CUAJUNGCO
• 金额: $ 13.4万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10287961
• 关键词: Affect; Affinity; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Binding; Biochemical; Biological; Biological Assay; Biological Phenomena; Biological Process; Brain; Brain region; Carrier Proteins; Cations; Cell membrane; Cells; Cerebrum; Chelating Agents; Communication; Cytoplasm; Data; Disease; Drug or chemical Tissue Distribution; Dyes; Event; Exhibits; Female; Fibroblasts; Financial compensation; FluoZin-3; Fluorescence; Fluorescence Microscopy; Ganglioside Sialidase Deficiency Disease; Genes; Genetic; Genotype; Glutamates; Goals; Health; Heterodimerization; Homeostasis; Human; Human Biology; Inductively Coupled Plasma Mass Spectrometry; Ion Channel; Isotopes; Knock-out; Knockout Mice; Knowledge; Laboratories; Left; Link; Lysosomes; Maintenance; Mammalian Cell; Metalloproteins; Metals; Molecular; Molecular Weight; Mus; Neurons; Observational Study; Organism; Parkinson Disease; Patients; Pattern; Phenotype; Physiology; Process; Property; Protein Family; Protein Isoforms; Proteins; Rattus; Regulation; Reporting; Rodent; Role; SLC30A3 gene; Sex Differences; Stains; Synaptic Vesicles; Synaptosomes; Techniques; Time; Tissues; Trace Elements; Trace metal; Validation; Vesicle; Western Blotting; Zebrafish; Zinc; brain tissue; cell type; differential expression; dimer; functional loss; human disease; insight; loss of function mutation; male; mind control; negative affect; pain perception; protein function; protein protein interaction; public health relevance; receptor; sex; silver sulfide; transcriptome; transcriptome sequencing; zinc-binding protein

PROJECT SUMMARY Zinc homeostasis in many organisms, including humans, is achieved by tissue-specific and highly conserved low molecular weight proteins known as the zinc transporter (ZnT) and Zrt- and Irt-like protein (ZIP) families. Several redundant ZnT and ZIP transporters have been identified in tissues that confer zinc efflux and influx property, respectively. A strict regulation of intracellular zinc levels exists in many tissues, especially in the brain, because this tissue contains a sizeable amount of chelatable zinc pool that is co- released with glutamate during normal neuronal communication. Indeed, the ZnT3 zinc effluxer has been shown to be important in sequestering and shuttling zinc into glutamatergic vesicles and that knocking out this protein obliterates vesicular zinc compartmentalization in the brain. We identified Transmembrane 163 (TMEM163), a zinc-binding protein and transporter, as a protein interactor of the TRPML1 ion channel. Loss of TRPML1 function causes Mucolipidosis IV disease. Meanwhile, TMEM163 was recently reported to modulate pain perception via its interaction with the neuronal P2X3 receptor ion channel. TMEM163 also known as synaptic vesicle 31 (SV31) protein was first identified in rat brain synaptosomes and exists as a dimer. TMEM163 localizes within the plasma membrane and vesicular compartments such as synaptic vesicles and lysosomes. We have preliminary evidence that TMEM163 interacts with the ZnT3 zinc efflux transporters. Functional zinc flux assays show that the efflux activity of TMEM163-ZnT3 heterodimers parallels that of their respective homodimer isoforms. These results not only confirm that TMEM163 is a zinc effluxer, but that its heterodimerization with a related zinc transporter adds to a repertoire of homeostatic control for intracellular zinc levels. Thus, it appears that TMEM163 is important for the maintenance of brain zinc homeostasis that is independent of, or in conjunction with, another zinc efflux transporter. The overarching goal of this project is to investigate the biological significance of human TMEM163 using its mouse Tmem163 counterpart. To this end, we will use a Tmem163 knockout (KO) mouse to determine changes in chelatable zinc distribution patterns in the brain using histochemical, histo- fluorescence, and biochemical techniques. We will also examine the Tmem163 KO phenotype by analyzing the brain transcriptome using RNA sequencing to establish whether genetic compensation (a well-known biological phenomenon) by other zinc transporter genes results from the loss of Tmem163 function. Overall, this project could fill current gaps in knowledge on the biological function of TMEM163, and could provide insights on how cells or tissues devoid of TMEM163 impact human health or human disease processes where zinc dyshomeostasis has been implicated.

项目摘要 包括人类在内的许多生物中的锌稳态是通过组织特异性和高度实现的 保守的低分子量蛋白称为锌转运蛋白（ZNT）以及Zrt-和IRT样蛋白 （拉链）家庭。在赋予锌的组织中已经鉴定出了几个冗余Znt和Zip转运蛋白 外排和涌入特性。严格对细胞内锌水平的调节存在于许多组织中， 尤其是在大脑中，因为该组织包含大量的可螯合锌池 在正常神经元通信期间用谷氨酸释放。确实，Znt3锌外排是 证明在隔离和将锌穿梭到谷氨酸囊泡中很重要，并且撞倒 该蛋白会抑制大脑中的水泡锌分区化。我们确定了跨膜163 （TMEM163），一种锌结合蛋白和转运蛋白，作为TRPML1离子通道的蛋白质相互作用。 TRPML1功能的丧失会导致粘脂脂异常IV疾病。同时，最近报道了TMEM163 通过与神经元P2X3受体离子通道的相互作用来调节疼痛感知。 TMEM163 也称为突触囊泡31（SV31）蛋白首先在大鼠脑突触体中鉴定出来并存在 作为二聚体。 TMEM163位于质膜和小囊室内的TMEM163 突触囊泡和溶酶体。我们有初步证据表明TMEM163与Znt3相互作用 锌外排转运蛋白。功能性锌通量测定表明TMEM163-ZNT3的外排活动 异二聚体与它们各自的同二聚体同工型的相似。这些结果不仅确认 TMEM163是一个锌外排，但与相关锌转运蛋白的异二聚化增加了 对细胞内锌水平的体内平衡控制的曲目。因此，TMEM163似乎很重要 为了维持与另一种锌独立或结合的脑锌稳态的 外排转运蛋白。该项目的总体目标是研究人类的生物学意义 TMEM163使用其鼠标TMEM163对应物。为此，我们将使用TMEM163淘汰赛（KO） 小鼠使用组织化学，组织 - 荧光和生化技术。我们还将通过 使用RNA测序分析脑转录组以确定遗传补偿是否（a） 其他锌转运蛋白基因的众所周知的生物学现象是由TMEM163的丧失产生的 功能。总体而言，该项目可以填补有关TMEM163的生物学功能知识的当前空白， 并可以提供有关细胞或组织如何影响人类健康或人类的见解 涉及锌dyshomeostisis的疾病过程。