Lipid Mediators and Lipid Metaborome

1溶血磷脂酰基转移酶的功能分析：通过In Silico数据库分析，我们鉴定了参与膜脂类生物合成重塑途径的几种溶血磷脂酰基转移酶。基于这些信息，我们成功地克隆了Lyso-PAF乙酰转移酶和LPCAT1。LYSO-PAF乙酰基转移酶是刺激依赖的PAR形成的关键酶，LPCAT1是参与肺表面活性物质中磷脂酰胆碱合成的乙酰基转移酶，在呼吸生理中起着关键作用。我们正在培育这些酶的基因缺陷小鼠，以阐明其生理重要性。2.磷脂酶A_2的功能分析：在许多哺乳动物中，胞浆磷脂酶A_2α(CPLA_2α)通过产生脂质介质在多种生理和病理生理条件下发挥重要作用。我们新发现了cpla_2家族酶，cpla_2δ，cpla_2ε和cpla_2…更多的ζ。我们正在分析这些酶的生理功能。3新的脂质介质受体(G蛋白偶联受体：GPCRs)的鉴定；寻找新的脂介质受体。150个孤儿的GPCR，我们成功地选择了大约。30个候选GPCR。在这些GPCRs中，我们新鉴定了TDAG8和G2A为质子敏感受体，LPA4为溶血磷脂酸受体。我们正在继续进行其他孤儿GPCRs的配基筛选。4对GPCR缺陷小鼠的分析；为了阐明脂质中介GPCRs的生理意义，我们产生了BLT1、BLT2、CysLT2和LPA4的基因缺陷小鼠。在对BLT1基因缺陷小鼠的分析中，我们获得了BLT1对于OVA诱导的哮喘模型中Th2细胞因子的产生和气道高反应性的发展所必需的证据。5通过LC-MS高通量定量脂质介质；通过结合脂层析和串联质谱仪(MS/MS)，我们开发了一种高度特异和合理灵敏的分析方法，在10分钟的单次运行中同时定量14种不同的介质。较少

1 Functional analysis of lysophospholipid acyltransferase ; We identified several lysophospholipid acyltransferases involving in the remodeling pathway of membrane lipids biosynthesis by in silico database analyses. Based on these information, we succeeded in molecular cloning of lyso-PAF acetyltransferase and LPCAT1. Lyso-PAF acetyltransferase is a critical enzyme in stimulus dependent formation of PAR LPCAT1 is an acetyltransferase involving in synthesis of phosphatidylcholine in pulmonary surfactant and play a pivotal role in respiratory physiology. We are generating gene-deficient mice of these enzymes to clarify the physiological importance. We further analyze other enzymes in this family.2 Functional analysis of phospholipase A_2 (PLA_2) ; Among many mammalian PLA_2s, cytosolic PLA_2 α(cPLA_2α) plays a critical role in various physiological and pathophysiological conditions through generating lipid mediators. We newly discovered cPLA_2 family enzymes, cPLA_2δ, cPLA_2ε, and cPLA_2 … More ζ . We are analyzing the physiological function of these enzymes.3 Identification of novel lipid mediator receptors (G-protein coupled receptors : GPCRs) ; To search for novel lipid mediator receptors among approx. 150 orphan GPCRs, we succeeded in selecting approx. 30 candidate GPCRs. In these GPCRs, we have newly identified TDAG8 and G2A as proton-sensing receptors and LPA4 as a lysophosphatidic acid receptor. We are continuously performing the ligand screening for other orphan GPCRs.4 Analysisi of GPCR-deficient mice ; To clarify the physiological significance of lipid mediator GPCRs, we have generated genedeficient mice for BLT1, BLT2, CysLT2 and LPA4. In the analysis of BLT1-deficient mice, we obtained evidence that BLT1 is necessary for production of Th2 cytokines and the development of airway hyperresponsiveness in an OVA-induced bronchial asthma model.5 High throughput quantitation of lipid mediators by LC-MS ; By the combination of lipid chromatography and tandem mass spectrometry (MS/MS), we developed a highly specific and reasonably sensitive assay procedure to simultaneously quantitate > 14 different mediators in 10 min single run. Less