权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Regulation of mitochondrial heme metabolism by Tmem14c

Tmem14c 对线粒体血红素代谢的调节

基本信息

批准号：
8677578
负责人：
Yvette Y Yien
金额：
$ 5.6万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-30 至 2015-06-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8677578
关键词：
Acute Erythroblastic Leukemia Affinity Chromatography Anemia Animal Model Biology Carrier Proteins Cell Culture Techniques Cells Child Complex Data Defect Development Disease Drug Metabolic Detoxication Environment Enzymes Erythrocytes Erythroid Erythroid Cells Erythropoiesis Foundations Genes Genetic Goals Health Hematological Disease Hematopoiesis Hematopoietic Heme Heme Iron Hemoglobin Human Iron Label Light Mammalian Cell Mass Spectrum Analysis Mediating Mentors Metabolic Pathway Metabolism Mitochondria Mitochondrial Proteins Mus Organ Oxidation-Reduction Oxygen Pathway interactions Patients Phenotype Play Porphyrias Porphyrins Process Production Pronormoblasts Prosthesis Proteins Public Health Reaction Regulation Research Research Infrastructure Respiration Role Staging Techniques Testing Thin Layer Chromatography Training Uroporphyrinogen III Vertebrates Woman Work Zebrafish coproporphyrinogen III deuteroporphyrin-IX embryonic stem cell erythroid differentiation iron metabolism novel oxygen transport porphyrin metabolism programs protoporphyrin IX public health relevance small hairpin RNA success tool trafficking

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to identify and characterize novel proteins that regulate mitochondrial heme metabolism, particularly proteins that play a role in mitochondrial heme/porphyrin transport. This project has implications for identifying genetic modifiers for erythroid porphyria and anemia, and thus has public health significance. Our lab has identified Tmem14c as a novel mitochondrial protein that is required for terminal erythroid differentiation and hemoglobinization in vertebrates. Heme synthesis enzymes are present in Tmem14c-deficient murine erythroleukemia (MEL) cells, and preliminary data from our lab indicate that iron transport in Tmem14c-deficient MEL cells is comparable to wild-type MEL cells. However, Tmem14c-deficient MEL cells synthesize decreased levels of heme and protoporphyrin IX. These data suggest that Tmem14c may facilitate mitochondrial porphyrin transport, thereby playing a key role in heme/porphyrin metabolism. In Aim 1, we will test this hypothesis by identifying the block in porphyrin metabolism in Tmem14c deficient cells, comparing the levels of uroporphyrinogen III (cytoplasmic), coproporphyrinogen III (mitochondrial/cytoplasmic) and protoporphyrin IX (mitochondrial) in the mitochondrial and cytoplasmic fractions of wild-type and Tmem14c-deficient MEL cells. In this manner, we will determine if the block in heme synthesis in Tmem14c-deficient cells is a result of defective porphyrin trafficking. As Tmem14c forms higher-order complexes with other proteins, we hypothesize that Tmem14c's interactions with its partners play a crucial role in regulating its function. In Aim 2, we will identify Tmem14c partner and characterize their role in regulating Tmem14c in the heme synthetic pathway; we will identify interacting proteins that are required for terminal erythroid differentiation, heme synthesis and porphyrin transport, and focus on characterizing proteins that regulate Tmem14c's localization and protein stability. Collectively, our studies will contribute to our lon-term objective of understanding regulatory mechanisms controlling heme metabolism, and the interaction of heme metabolism with erythropoiesis. The specific aims in this proposal are of particular significance because studies of Tmem14c function will shed light on the poorly understood porphyrin trafficking pathways that are central to the regulation of heme metabolism and erythropoiesis. These experimental aims are a logical continuation of my graduate work in hematopoiesis, but provide a framework with which to obtain substantial training in the use of the zebrafish as an animal model and additional techniques that will prove invaluable for further studies in erythroid biology.

描述（由申请人提供）：本项目的长期目标是鉴定和表征调节线粒体血红素代谢的新型蛋白质，特别是在线粒体血红素/卟啉转运中发挥作用的蛋白质。该项目对鉴定红系卟啉病和贫血的遗传修饰剂具有意义，因此具有公共卫生意义。我们的实验室已经确定Tmem 14 c是一种新的线粒体蛋白，是脊椎动物终末红细胞分化和血红蛋白化所必需的。血红素合成酶存在于Tmem 14 c缺陷型小鼠红白血病（MEL）细胞中，我们实验室的初步数据表明，Tmem 14 c缺陷型MEL细胞中的铁转运与野生型MEL细胞相当。然而，Tmem 14 c缺陷MEL细胞合成血红素和原卟啉IX的水平降低。这些数据表明，Tmem 14 c可能有助于线粒体卟啉转运，从而在血红素/卟啉代谢中发挥关键作用。在目的1中，我们将通过鉴定Tmem 14 c缺陷细胞中卟啉代谢的阻断，比较野生型和Tmem 14 c缺陷MEL细胞的线粒体和细胞质组分中尿卟啉原III（细胞质）、粪卟啉原III（线粒体/细胞质）和原卟啉IX（线粒体）的水平来测试这一假设。通过这种方式，我们将确定Tmem 14 c缺陷细胞中血红素合成的阻断是否是有缺陷的卟啉运输的结果。由于Tmem 14 c与其他蛋白质形成高阶复合物，我们假设Tmem 14 c与其伴侣的相互作用在调节其功能方面起着至关重要的作用。在目标2中，我们将确定Tmem 14 c的合作伙伴，并表征其在血红素合成途径中调节Tmem 14 c的作用;我们将确定终末红细胞分化，血红素合成和卟啉转运所需的相互作用蛋白，并专注于表征调节Tmem 14 c的定位和蛋白质稳定性的蛋白质。总的来说，我们的研究将有助于我们了解控制血红素代谢的调节机制以及血红素代谢与红细胞生成的相互作用的长期目标。在这一建议的具体目标是特别重要的，因为Tmem 14 c功能的研究将阐明知之甚少的卟啉运输途径，是中央血红素代谢和红细胞生成的调节。这些实验目标是我的造血研究生工作的逻辑延续，但提供了一个框架，以获得大量的培训，在使用斑马鱼作为动物模型和额外的技术，将证明是非常宝贵的进一步研究红细胞生物学。