Mitochondrial Lipid Kinase

线粒体脂质激酶

• 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
• 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

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与鞘氨醇(SPHK1，2)和神经酰胺(CERK)激酶以及神经酰胺激酶样蛋白(CERKL)一起组成鞘磷脂激酶家族。AGK在这个家族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等位基因的小鼠由于植入失败而在胚胎发育早期死亡，这也不同于SPHKs、Cerk或CERKL，在这些地方，空小鼠是存活和可生育的。然而，AGK催化的磷酰化转移反应的脂质底物尚不确定。我们提出的研究计划将发现底物/产物，并在此过程中定义一条很可能对线粒体生存至关重要的脂代谢途径。具体地说，我们将：(1)建立AGK表达明显不同的细胞培养和组织的配对，并使用质谱学来表征这些细胞和组织的脂体，从而最终确定AGK催化的反应；(2)通过确定其亚细胞器位置，研究AGK缺乏细胞的线粒体生理学，并通过条件缺失AGK等位基因来确定细胞谱系的命运，来表征线粒体中AGK的功能。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，再加上线粒体生理学方面的专业知识，将使我们能够解决这个问题。至少，拟议中的实验将揭示鞘磷脂代谢的一个新分支。最大限度地，我们将定义一条与线粒体功能不可或缺的新途径。 与公共卫生相关：许多严重的病理学以身体能量工厂的缺陷为特征，这些工厂是称为线粒体的细胞器。根据线粒体疾病影响的器官系统，症状可能包括生长不良、肌肉无力、糖尿病、癫痫、呼吸障碍和痴呆症等神经问题。在这个项目中，我们研究了一种名为AGK的蛋白质，它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的小鼠在胚胎形成时早期死亡，这表明AGK对包括人类在内的哺乳动物的发育是必不可少的。我们正在研究AGK，以了解受AGK影响的脂质途径的性质，以及为什么这一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更好地了解、诊断并最终治疗线粒体疾病。