1991 Fiscal Year Final Research Report Summary
Study on the physiological and pathological dynamics of cyclooxygenase and lipoxygenase enzymes
| Project/Area Number |
01440025
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
YAMAMOTO Shozo Department of Biochemistry, Tokushima University, School of Medicine, Professor, 医学部, 教授 (50025607)
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| Co-Investigator(Kenkyū-buntansha) |
UEDA Natsuo Department of Biochemistry, Tokushima University, School of Medicine, Lecturer, 医学部, 講師 (20193807)
TAKAHASHI Yoshitaka Department of Biochemistry, Tokushima University, School of Medicine, Instructor, 医学部, 助手 (10236333)
HAYASHI Yoko Department of Biochemistry, Tokushima University, School of Medicine, Instructor, 医学部, 助手 (60035441)
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| Project Period (FY) |
1989 – 1991
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| Keywords | Lipoxygenase / Cyclooxygenase / Arachidonic acid / Prostaglandin / Leukotriene / Monoclonal antibody / cDNA / Enzyme immunoassay |
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| Research Abstract |
Bioactive compounds like prostaglandin, thromboxane and leukotriene are synthesized from arachidonic acid by cyclooxygenase and lipoxygenase. This research project is to investigate the physiological and pathological dynamics of these enzymes at cellular and molecular levels. The followings were our approaches for this research-purpose. (1) Catalytic properties of the enzymes : 12-Lipoxygenase of human platelets was immunoaffinity-purified, and its substrate specificity was studied. (2) Enzyme distribution screened by peroxidaselinked immunoassay : 12-Lipoxygenase was detected in a considerable quantity in porcine anterior pituitary. (3) Enzyme distribution screened by immunohistochemical methods : Among porcine tissues 5- and 12-lipoxygenases were localized in granulocytes but not in lymphocytes. In addition, 5-lipoxygenase was found in lung and pancrease, and 12-lipoxygenase in pituitary. (4) Assay of cyclooxygenase metabolites : A monoclonal antibody against 11-dehydro-thromboxane B2 was raised. and utilized for its inununoaffinity purification and immunoassay. (5) Physiological anc pathological dynamics of enzymes : Cyclooxygenase induction in mouse osteoblastic cells by EGF, TGF or epinephrine was confirmed by Western blotting using anti-cyclooxygenase antibody and by Northern blotting using cyclooxygense cDNA. (6) cDNA cloning : We cloned cDNAs for 12-lipoxygenases of porcine leukocytes and human erythroleukemia cells and cyclooxygenase of human platelets, and the primary structures of these enzymes were deduced from the nucleotide sequences of these cDNAs.
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[Publications] Maruyama, T., Ueda, N., Yoshimoto, T., Yamamoto, S., Komatsu, N., and Watanabe, K.: "Immunohistochemical study of arachidonate 12-lipoxygenase in porcine tissues" J. Histochem. Cytochem.37. 1125-1131 (1989)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Ueda, N., Hiroshima, A., Natsui, K., Shinjo, F., Yoshimoto, T., Yamamoto, S., Ii, K., Gerozissis, K., and Dray, F.: "Localization of arachidonate 12-lipoxygenase in parenchymal cells of porcine anterior pituitary" J. Biol. Chem.265. 2311-2316 (1990)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Hayashi, Y., Shono, F., Yamamoto, S., Takasaki, W., Nakagawa, A., Watanabe, K., Yamashita, K., and Miyazaki, H.: "Immunoaffinity purification of 11-dehydro-thromboxane B_2 from human urine and plasma for quantitative analysis by radioimmunoassay" Anal. Biochem.187. 151-159 (1990)
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「研究成果報告書概要(欧文)」より
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[Publications] Yoshimoto, T., Suzuki, H., Yamamoto, S., Takai, T., Yokoyama, C., and Tanabe, T.: "Cloning and sequence analysis of the cDNA for arachidonate 12-lipoxygenase of porcine leukocytes" Proc. Natl. Acad. Sci. USA. 87. 2142-2146 (1990)
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「研究成果報告書概要(欧文)」より
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[Publications] Oshima, T., Yoshimoto, T., Yamamoto, S., Kumegawa, M., Yokoyama, C., and Tanabe, T.: "cAMP-dependent induction of fatty acid cyclooxygenase mRNA in mouse osteoblastic cells (MC3T3-E1)" J. Biol. Chem.266. 13621-13626 (1991)
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