Biochemical Study on Leukoriene D_4 Receptor in the Lung

本课题的目的是对白三烯（LT） D_4或B_4受体进行分子克隆或纯化，从分子水平上了解白三烯的信号转导机制。我们首次用一种洗涤剂（CHAPS）将豚鼠肺中的LTD_4受体和猪脾脏中的LTB_4受体以活性形式溶解。使用这种可溶解的制剂，我们可以部分纯化受体，并表征受体蛋白及其相关的gtp结合蛋白（参考文献1,2）。利用豚鼠肺mRNA构建cDNA文库。利用zenopus卵母细胞表达系统，我们试图从该文库中获得一个编码LTD_4受体和血小板活化因子（PAF）受体的克隆。我们仍在尝试LTD_4受体，但已经得到了一个克隆编码PAF受体。我们明确了受体蛋白的结构，并成功地在哺乳动物（COS）细胞中表达了该受体。

The objective of this project is the molecular cloning or purification of leukotriene (LT) D_4 or B_4 receptor for the understanding of signal transduction mechanism of leukotiene at the molecular level. We solubilized LTD_4 receptor in an active form from guinea pig lung and LTB_4 receptor from porcine spleen using a detergent (CHAPS) for the first tine. Using this solubilized preparation, we could partially purify the receptor and characterize the receptor protein and its associated GTP-binding protein (Ref. 1,2). We constructed cDNA library from guinea pig lung mRNA. Using zenopus oocyte expression system, we tried to get a clone encoding LTD_4 receptor and also Platelet-Activating Factor(PAF) receptor from this library. We are still trying for LTD_4 receptor, but have gotten a clone encoding PAF receptor. We clarified the structure of the receptor protein and also succeeded to express the receptor in a mammalian (COS) cells.