1990 Fiscal Year Final Research Report Summary
Biochemical Study on Leukoriene D_4 Receptor in the Lung
| Project/Area Number |
01570125
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | University of Tokyo |
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Principal Investigator |
WATANABE Tsuyoshi Assistant Professor Dept. of Int. Med., University of Tokyo, 医学部(病)・第一内科, 助手 (80158641)
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| Co-Investigator(Kenkyū-buntansha) |
TERAMOTO Tamio Assistant Professor, Dept. of Int. Med., University of Tokyo, 医学部(病)第一内科, 助手 (20133077)
SHIMIZU Takao Associate Professor, Dept. of Nutrition and Physiological Chemistry, University, 医学部栄養学, 助教授 (80127092)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Leukoriene / Lung / Receptor / Solubilization / Pertussis Toxin / Molecular Cloning / Oocytes / GTP-binding protein |
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| Research Abstract |
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.
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[Publications] Z. Honda, M. Nakamura, I. Miki, M. Minami, T. Watanabe, Y. Seyama, H. Okado, H. Toh, T. Miyamoto, and K. Ito, T. Shimizu: "Cloning by functional expression of guinea pig lung platelet activating factor (PAF) receptor" Nature. 349. 342-346 (1991)
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