Mitochondrial Lipid Kinase

线粒体脂质激酶

基本信息

批准号：
8241280
负责人：
KEVIN R. LYNCH
金额：
$ 19.11万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-10 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8241280
关键词：
Affect Alcohols Alleles Biochemical Biological Assay Biology Cell Lineage Cell physiology Cells Cellular biology Ceramides Chemicals Complement Coupled Data Defect Dementia Development Diabetes Mellitus Diacylglycerol Kinase Diagnosis Embryo Embryonic Development Endoplasmic Reticulum Enzymes Failure Family Family member Growth Human Implant Incubated Inner mitochondrial membrane Intention Knockout Mice Knowledge Laboratories Learning Lipids Location Lysophospholipids Mammals Mass Spectrum Analysis Membrane Metabolic Pathway Metabolism Mitochondria Mitochondrial Diseases Mitochondrial Matrix Mitochondrial Proteins Mus Muscle Weakness Names Nature Neurologic Organelles Pathology Pathway interactions Phenotype Phosphotransferases Physiology Problem Solving Proteins RNA Interference Reaction Recombinants Research Research Personnel Respiration Disorders Seizures Sphingolipids Sphingosine Staging Symptoms T-Lymphocyte Techniques Testing Tissues Work base body system ceramide kinase experience implantation lipid metabolism preimplantation programs research study sphingosine kinase tissue/cell culture tool

项目摘要

DESCRIPTION (provided by applicant): AGK, along with the sphingosine (SPHK1, 2) and ceramide (CERK) kinases and the ceramide kinase like protein (CERKL), comprise the sphingolipid kinase family. AGK is unique among members of this family in that it is a mitochondrial protein. We discovered that mice lacking a functional AGK allele die early in embryogenesis due to failure to implant, which is also unlike SPHKs, CERK or CERKL where null mice are viable and fertile. However, the lipid substrate of phosphoryl transfer reaction catalyzed by AGK is uncertain. The research program we propose will discover that substrate/product and in doing so will define a lipid metabolic pathway that is most likely crucial to mitochondrial survival. Specifically, we will: (Aim 1) Generate matched pairs of cell cultures and tissues wherein AGK expression is markedly different and use mass spectrometry to characterize the lipidome of those cells and tissues so as to ultimately identify the reaction catalyzed by AGK and (Aim 2) characterize AGK function in mitochondria by determining its sub- organelle location, studying mitochondrial physiology in cells deficient in AGK and using conditional deletion of AGK alleles to determine the fate of cell lineages in the mouse. Our extensive experience studying lysophospholipid chemical biology including sphingosine kinases coupled with expertise in mitochondrial physiology will enable us to solve this problem. Minimally, the experiments proposed will reveal a new branch of sphingolipid metabolism. Maximally, we will define a new pathway that is integral to mitochondrial function. PUBLIC HEALTH RELEVANCE: A number of serious pathologies are characterized by defects in the body's energy factories, which are cell organelles named mitochondria. Depending on the organ system affected in mitochondrial diseases, symptoms might include poor growth, muscle weakness, diabetes, neurological problems such as seizures, respiratory disorders and dementia. In this project, we study a protein named AGK, which is thought to be involved in lipid metabolism. AGK is a mitochondrial protein and we discovered that mice lacking this protein die early in embryo formation, which indicates that AGK is essential for development in mammals including humans. We are researching AGK to learn the nature of the lipid pathway that is affected by AGK and why this pathway is so important to mitochondrial survival. Our intention is to use this knowledge to provide better understanding, diagnosis and, eventually, treatment of mitochondrial disease.

描述（由申请人提供）：AGK，以及鞘氨酸（SPHK1、2）和神经酰胺（Cerk）激酶以及蛋白质（CERKL）的神经酰胺激酶，包括鞘脂激酶家族。 AGK在这个家族的成员中是独一无二的，因为它是一种线粒体蛋白。我们发现缺乏功能性AGK等位基因的小鼠在胚胎发生早期死亡，因为未植入植入物，这也与无效小鼠可行且肥沃的SPHKS，CERK或CERKL不同。然而，尚不确定由AGK催化的磷酸化转移反应的脂质底物。我们提出的研究计划将发现底物/产物并这样做将定义一种脂质代谢途径，该途径最有可能对线粒体生存至关重要。具体而言，我们将：（ AIM 1）生成匹配的细胞培养物和组织对，其中AGK表达明显不同，并使用质谱法来表征这些细胞和组织的脂质组，从而最终识别由AGK催化的反应和AIM 2（AIM 2））通过确定其亚细胞器位置，研究AGK功能在AGK缺乏的细胞中的线粒体生理以及使用AGK等位基因的条件缺失来确定小鼠细胞谱系的命运。我们研究溶血磷脂化学生物学的丰富经验，包括鞘氨醇激酶以及线粒体生理学专业知识，将使我们能够解决这个问题。最少，提出的实验将揭示鞘脂代谢的新分支。最大程度地，我们将定义一种与线粒体功能不可或缺的新途径。公共卫生相关性：许多严重的病理的特征是人体能量工厂的缺陷，这些病情是线粒体的细胞细胞器。根据线粒体疾病受影响的器官系统，症状可能包括生长不良，肌肉无力，糖尿病，神经系统问题，例如癫痫发作，呼吸系统疾病和痴呆症。在这个项目中，我们研究了一种名为AGK的蛋白质，该蛋白质被认为与脂质代谢有关。 AGK是一种线粒体蛋白，我们发现缺乏这种蛋白质的小鼠在胚胎形成早期死亡，这表明AGK对于包括人类在内的哺乳动物的发育至关重要。我们正在研究AGK，以学习受AGK影响的脂质途径的性质，以及为什么该途径对线粒体生存如此重要。我们的目的是利用这些知识来提供更好的理解，诊断以及最终对线粒体疾病的治疗。