Biochemical Study on Leukoriene D_4 Receptor in the Lung

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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.