薬への応用を目差したアラキドン酸代謝制御とそれによる細胞情報伝達の生化学的解明

1990 年度 実績報告書

• 研究課題/領域番号: 01304049
• 研究機関: 京都大学
• 研究代表者: 市川 厚 京都大学, 薬学部, 教授 (10025695)
• 研究分担者: 山本 尚三 徳島大学, 医学部, 教授 (50025607) ; 奥山 治美 名古屋市立大学, 薬学部, 教授 (90080176) ; 成宮 周 京都大学, 医学部, 助教授 (70144350) ; 名取 俊二 東京大学, 薬学部, 教授 (50012662) ; 井上 圭三 東京大学, 薬学部, 教授 (30072937)
• キーワード: アラキドン酸代謝 / プロスタグランジン / リポキシゲナ-ゼ / レセプタ- / αーリノレン酸 / 細胞情報伝達 / ロイコトリエン

研究概要

本年度の研究計画に従い以下の研究成果を得た。
1.アラキドン酸代謝系の開始に係わる細胞情報伝達とアラキドン酸生成系の制御に関する研究;
(1)アラキドン酸遊離に関与する炎症性ホスホリパ-ゼA_2(14kDa)の一次構造を解析し、その阻害タンパク質(C3産物)を発見(井上)。(2)血小板のアラキドン酸代謝を制御するPGE_1はG蛋白smgp21をリン酸化するがそれはAキナ-ゼ部位と同一であることを発見(高井)。(3)3T3細胞の血清依存的増殖活性を停止すると細胞膜G蛋白は増加し新たな核局在G蛋白(140kDa)が減少することを発見した(名取)。(4)ラット好中球のPAF産生、およびマウスマクロファ-ジからのTNF産生がαーリノレン酸で抑制されることを発見した(奥山)。(5)白血球のO_2^ー産生を行うNADPH oxidase活性の制御因子と、アラキドン酸による因子の活性化(細胞質→細胞膜変換)を発見(石橋)。
2.アラキドン酸代謝産物の生合成制御とその修飾に関する研究;
(1)精製した5ーリポキシゲナ-ゼを用い、抗アレルギ-効果をもつフラボノイド(Cirsillol)が酵素を選択的に阻害するのを発見(山本)。(2)5ーリポキシゲナ-ゼ阻害と抗ヒスタミン作用を併せもつ抗アレルギ-薬を検索し、経口投与可能なTMKー688を開発した(室田)。
3.アラキドン酸代謝産物の作用機構の解析に関する研究;
(1)ヒト血小板のトロンボキサンA_2受容体を単離し、cDNAから受容体の一次構造を決定(成宮)。(2)モルモット肺の細胞膜からLTD_4受容体を可溶化し諸性質を解析した。また、PAF受容体のcDNAクロ-ニングを行い一次構造を決定(清水)。(3)癌化肥満細胞のPGD_2受容体のリン酸化反応、PGE_1受容体の糖鎖付加による細胞膜移行、PGI_2/PGE_2受容体の情報伝達系を解析した(市川)。(4)トロンビン刺激肥満細胞におけるアラキドン酸代謝やCa^<2+>代謝のdown regulationに働く細胞膜G蛋白の細胞質移行反応を発見(市川)。

研究成果

• [文献書誌] Kimio Yatsunami: "Effect of tunicamycin on functions of PGE_1 receptors from mouse mastocytoma Pー815 cells" Biochimica et Biophysica Acta. 1051. 94-99 (1990)
• [文献書誌] Hitoshi Hashimoto: "Identification of a prostacyclin receptor copled to the adenylate cyclase system via a stimulatory GTPーbinding protein in mouse mastocytoma Pー815 cells" Prostaglandins. 40. 491-505 (1990)
• [文献書誌] K.Koizumi: "Induction of 31,000 protein by PGD_2ーtreated vascular endothelial cells" Advances in Prostaglandin,thromboxane,and Leukotriene Research. 21. 891-894 (1990)
• [文献書誌] Yasuhiro Hayashi: "Hypoxanthine increases basal and FGFーstimulated growth of porcine aortic endothelial cells in media with calf of newborn bovine serum" In Vitro Cell.Dev.Biol.26. 843-845 (1990)
• [文献書誌] Jun Yamamoto: "cDNAーderived amino acid sequence of Lーhistidine decarboxylase from mouse mastocytoma Pー815 cells" FEBS LETTERS. 276. 214-218 (1990)
• [文献書誌] Kimio Yatsunami: "Eicosanoids and their receptors" Seikagaku. 62. 343-348 (1990)
• [文献書誌] Toshiaki Ohtsuka: "A diacylglycerol kinase inhibitor,R59022,potentiates superoxide anion production and 46ーkDa protein phosphorylation in guinea pig polymorphonuclear leukocytes" The Journal of Biological Chemistry. 265. 15418-15423 (1990)
• [文献書誌] Shozo Yamamoto: "Flavonoids as 5ーlipoxygenase inhibitors" Flavonoides in Biology & Medicine III. (1990)
• [文献書誌] Natsuo Ueda: "The 6Rーoxygenase activity of arachidonate 5ーlipoxygenase purified from porcine leukocytes" The Journal of Biological Chemistry. 263. 1937-1941 (1988)
• [文献書誌] Tanihiro Yoshimoto: "Cloning and sequence analysis of the cDNA for arachidonare 12ーlipoxygenase of porcine leukocytes" Proc.Natl.Acad.Sci.USA. 87. 2142-2146 (1990)
• [文献書誌] S.Murota: "Development of an orally active antiallergic drug: TMKー688" New Trends in lipid Mediators Research. 3. 56-61 (1989)
• [文献書誌] Yoshiharu Suzuki: "Cadmium stimulates prostaglandin E_2 synthesis in osteoblastーlike cells,MC3T3ーE1" Biochimica et Biophysica Acta. 1021. 135-139 (1989)
• [文献書誌] Kazuhiro Toriyama: "Prostaglandin E_2 evokes intracellular calcium rise in mouse osteoblastic cell line,MC3T3Eー1" Eicosanoids. 3. 157-160 (1990)
• [文献書誌] Masakazu Hirata: "Cloning and expression of cDNA for a human thromboxane A_2 receptor" Nature. 349. 617-620 (1991)
• [文献書誌] Nobuya Ohishi: "Immunological quantitation and immunohistochemical localization of leukotriene A_4 hydrolase in guinea pig tissues" The Journal of Biological Chemistry. 265. 7520-7525 (1990)
• [文献書誌] Tsuyoshi Watanabe: "Characterization of the guinea pig lung membrane leukotriene D_4 receptor solubilized in an active form" The Journal of Biological Chemistry. 265. 21237-21241 (1990)
• [文献書誌] Masayuki Komada: "Structure of gene coding for rat group II phospholipase A_2" Biochemical and Biophysical Research Communications. 168. 1059-1065 (1990)
• [文献書誌] Yorimasa Suwa: "Proteinaceous inhibitors of phospholipase A_2 purified from inflammatory sites in rats" Proc.Natl.Acad.Sci.USA. 87. 2395-2399 (1990)
• [文献書誌] Dae Kyong Kim: "Detection and subcellular localization of rabbit platelet phospholipase A_2 which preferentially hydrolyzes an arachidonoyl residue" J.Biochem.108. 903-906 (1990)
• [文献書誌] Kana Kamano: "Effects of a highーlinolate and a highーαーlinolenate diet on spontaneous mammary tumourigenesis in mice" Anticancer Research. 9. 1903-1908 (1989)
• [文献書誌] Shiro Watanabe: "Effect of dietary αーlinolenate/linoleate balance on crescent typeーantiーglomerular basement membrane Nephritis in rats" Lipids. 25. 267-272 (1990)
• [文献書誌] Masao Hirose: "Effects of dietary perilla oil,soybean oil and safflower oil on 7,12ーdimethylbenz[α]anthracene(DMBA) and 1,2ーdimethylhydrazine(DMH)ーinduced mammary gland and colon carcinogenesis in femal SD rats" Carcinogenesis. 11. 731-735 (1990)
• [文献書誌] Masahiko Hoshijima: "Phosphorylation by cyclic AMPーdependent protein kinase of a human platelet mr 22,000 GTPーbinding protein (smg p21) having the same putative effector domain as the ras gene products" Biochemical and Biophysical Research Communications. 157. 851-860 (1988)
• [文献書誌] Masahito Kawata: "Phosphorylation of smg p21,a ras p21ーlike GTPーbinding protein,by cyclic AMPーdependent protein kinase in a cellーfree system and in response to prostaglandin E_1 in intact human platelets" The Journal of Biological Chemistry. 264. 15688-15695 (1989)
• [文献書誌] Shizu Takeda: "Nuclear GTPーbinding proteins of Swiss 3T3 cells" FEBS Letters. 244. 469-472 (1989)
• [文献書誌] Fukuichi Ohsawa: "Purification and properties of a βーgalactosideーbinding lectin from neonatal marmoset" J.Biochem.107. 431-434 (1990)
• [文献書誌] Masaki Ozawa: "Synergism between protein kinase C activator and fatty acids in stimulating superoxide anion production in guinea pig polymorphonuclear leukocytes" Archives of Biochemistry and Biophysics. 273. 491-496 (1989)
• [文献書誌] Toshiaki Ohtsuka: "Involvement of membrane charges in constituting the active form of NADPH oxidase in guinea pig polymorphonuclear leukocytes" Archives of Biochemistry and Biophysics. 280. 74-79 (1990)