Mitochondrial Lipid Kinase

线粒体脂质激酶

• 批准号: 8410575
• 负责人: KEVIN R. LYNCH
• 金额: $ 21.78万
• 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 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8410575
• 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.

描述（由申请人提供）：AGK与鞘磷脂（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等位基因的小鼠由于植入失败而在胚胎发生早期死亡，这也与sphk、CERK或CERKL不同，其中无基因小鼠是存活和可育的。然而，AGK催化的磷酸化酰基转移反应的脂质底物是不确定的。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个脂质代谢途径，这可能对线粒体生存至关重要。具体而言，我们将：（目标1）生成匹配的细胞培养物和组织，其中AGK表达明显不同，并使用质谱法表征这些细胞和组织的脂质组，从而最终鉴定AGK催化的反应；（目标2）通过确定其亚细胞器位置来表征AGK在线粒体中的功能；研究AGK缺乏细胞的线粒体生理，并利用AGK等位基因的条件缺失来确定小鼠细胞系的命运。我们丰富的溶血磷脂化学生物学研究经验，包括鞘氨醇激酶，再加上线粒体生理学的专业知识，将使我们能够解决这个问题。至少，所提出的实验将揭示鞘脂代谢的一个新分支。最重要的是，我们将定义一个线粒体功能不可或缺的新途径。