EICOSANOID BIOSYNTHESIS DEFICIENCY

类二十烷酸生物合成缺陷

• 批准号: 7731407
• 负责人: JOHN Alexander OATES
• 金额: $ 0.02万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2007-09-16
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7731407
• 关键词: Anabolism; Biochemical; Calcium; Cells; Clinical; Complex; Computer Retrieval of Information on Scientific Projects Database; Defect; Eicosanoids; Eicosatetraenoic Acids; Enzymes; Funding; Genes; Genetic; Genotype; Grant; Growth Factor Oncogenes; Human; Individual; Institution; Knowledge; Lipoxygenase; Maps; Measures; Molecular; Molecular Genetics; Mutation; Pathway interactions; Patients; Phospholipase A2; Phospholipids; Phosphorylation; Platelet aggregation; Process; Prostaglandin-Endoperoxide Synthase; Prostaglandins; Protein Analysis; Regulation; Research; Research Personnel; Resources; Role; Serum; Source; United States National Institutes of Health; Western Blotting; cyclooxygenase 1; cyclooxygenase 2; cytokine; urinary

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Primary Hypothesis: The identified patient has a mutation of the cytosolic PLA2 enzyme. Secondary Hypothesis: The identified patient has a defect in regulatory mechanism of cytosolic PLA2. We have identified a patient with significantly low levels of multiple measured prostaglandins, products of the cyclooxygenase pathway, and 12-hydroxy-eicosatetraenoic acid (12-HETE), a product of the lipoxygenase pathway, suggesting a functional PLA2 enzymatic deficiency. We seek to further elucidate the molecular mechanism for these biochemical deficiencies. We will initially quantify serum and urinary metabolites of common cyclooxygenase and lipoxygenase pathways, in addition to functional analysis of platelet aggregation and phospholipid characterization to confirm the molecular defect in this individual. Following confirmation of the enzymatic deficiency, we will assess whether the enzyme is entirely absent from the cells or present in an altered form by performing Western blot analysis of the protein. We will then pursue description on a genetic level. PLA2 regulation in humans involves a complex process including calcium release-regulated activation, phosphorylation, translocation, and multiple other modulatory factors such as cytokines, growth factors, and oncogenes. Because human cytosolic and secretory PLA2 genes have been mapped and functional domains well described, genotyping of these enzymes will assist in pinpointing the mechanism of functional deficiency in our patient and delineating between a defect in the PLA2 enzyme itself or in a regulating mechanism. Defining the deficiency in this patient on molecular and genetic levels will contribute significantly to the knowledge of the role of PLA2 enzymes in humans and clinical consequences of their inhibition.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 主要假设：确定的患者具有胞质PLA2酶的突变。 次要假设：确定的患者在胞质PLA2的调节机制方面存在缺陷。 我们已经确定了一名患有多种测量的前列腺素，环氧合酶途径的产物和12-羟基 - 耐核酸乙烯烯酸（12-HETE）的患者，这是脂氧合酶途径的产物，这表明功能性PLA2酶降低。 我们寻求进一步阐明这些生化缺陷的分子机制。 除了对血小板聚集和磷脂表征的功能分析以确认该个体中的分子缺陷外，我们最初还将定量常见环氧酶和脂氧酶途径的血清和尿代谢产物。 在确认酶缺乏症后，我们将通过对蛋白质的蛋白质印迹分析进行蛋白质印迹分析来评估该酶是否完全不存在于细胞中或以改变形式。 然后，我们将在遗传水平上进行描述。 人类中的PLA2调节涉及一个复杂的过程，包括钙释放的激活，磷酸化，易位以及多种其他调节因素，例如细胞因子，生长因子和癌基因。 由于已经映射了人类的胞质和分泌PLA2基因并进行了充分描述的功能结构域，因此这些酶的基因分型将有助于确定患者中功能不足的机制，并在PLA2酶本身或调节机制中的PLA2酶本身或PLA2酶的缺陷之间进行描述。 定义该患者在分子和遗传水平上的缺乏将显着促进PLA2酶在人类中的作用以及其抑制作用的临床后果。