1992 Fiscal Year Annual Research Report

励起分子の関与する精密構造認識と機能発現

Research Project

Project/Area Number	04220105
Research Institution	The University of Electro-Communications
Principal Investigator	大橋守電気通信大学, 電気通信学部, 教授 (70015535)
Co-Investigator(Kenkyū-buntansha)	辻フレデリック一朗大阪バイオサイエンス研究所, 第2研究部, 部長 (80201755) 中山仁北海道大学, 薬学部, 助手 (70088863) 斉藤烈京都大学, 工学部, 教授 (20026082) 磯部稔名古屋大学, 農学部, 教授 (00023466)
Keywords	生物発光 / 化学発光 / エクオリン / 月夜茸 / 発光蛋白質 / DNAの光切断 / カルシウムチャンネル / ナトリウムチャンネル
Research Abstract	ホタル・ウミホタル・ホタルイカ等の生物発光は生体反応の精密な構造認識の結果生成される励起分子の緩和過程の一つであり、一方光合成,視覚,光感作のような受光系は光を吸収して生じる励起分子の構造変化を伴う光エネルギーの生体信号への変換系である。本研究はこのような発光,受光系にみられる励起分子と蛋白質の構造認識を明らかにし、その機能発現の機構を解明することが目的である。本年度中に得られた成果を次にまとめる。発光グループ:発光蛋白質エクオリンに開し、大橋はアポエクオリ及び関連蛋白質のESI-MSスペクトルをうることに成功し、0.05%の精度で分子量21,640を検出した。又エクオリンの発光種が従来信じられていたセレンテラミドのアミドアニオン種でなく、フェノラート・イオンの励起分子であることを示した。辻らは部位特異的変異法によりアポエクオリンの169番目のヒスチジン残基が酵素活性の中心部位である可能性を示唆した。一方磯部はトビイカの発光器からセレンテラミンを抽出同定し、抽出液の化学発光からルシフェリンはセレンテラジンであることを示唆した。又月夜茸の発光体はランブテロフラビンでありその発光に必要な三成分の一つがチロシンであることを確認した。受光グループ:斉藤はDNAの特定塩基配列を認識し、光励起種により特異的にDNAを切断する分子の開発を行い、アデニン-ブロモウラシルの配列で特異的にDNAを光切断する手法を見出し、その切断機構を分子レベルで明らかにした。一方中山はフォトアフィニテイラベル法に〔 ^3H〕ジアジピンを用い、心筋Caチャンネルのジヒドロピリジン結合部位を同定した。またNaチャンネルのテトロドトキシン結合部位も光アフィニテイラベル法で解明し、ドメインI,II,IIIについて新知見を得た。

Research Products

(22 results)

All Other

All Publications (22 results)

[Publications] H.Nakayam,Y.Hatanaka,E.Yoshida,K.Oka,Y.Kanaoka et al.: "Photolabeled sites with a tetrodotosin dericative in the domain III and Iv of the electroplax sodium channel" Biochem.Biophys.Res.Commun.184. 900-907 (1992)
[Publications] Y.Hatanaka,H.Nakayama,Y.Kanaoka: "Photoaffinity labcling of the electroplax sodium channel with a photorcactive μ-conotoxin carring a radioactive and chromogenic derivative." Chem.Pharm.Bull.40. 2597-2539 (1992)
[Publications] A.Kuniyasu,K.Oka,T.Ide-Yamada,Y.,Hatanaka,H.Nakayama et al: "Structural characterization of the cardiac calcium channel from procine hearts" J.Biochem.112. 235-242 (1992)
[Publications] H.Nakayam,A.Yamamoto,Y.Hatanaka,T.Hachisu,S.Tsunasawa et al.: "Characterization of saccharide moiety in the electroplax sodium channel" Chem.Pharm.Bull.40. 3287-3291 (1992)
[Publications] H.Nakayama,H.Shikano,Y.Kanaoka: "A site directed antibody which inhibits phosphorylation of the rat brain sodium channel by CAMP-dependent protein kinase" Biochem.Biophys.Acta. 1175. 67-72 (1992)
[Publications] F.Q.Chen,T.Hirano,M.Ohashi,H.Nakayama,K.Oda,M.Machida: "3-Benzoy1-5-(4 hydroxyphenyl)-2-[4-(1-azi-2,2,2-trifluoroethyl)phenylacetanido]phrazine.A photosensitive analogue of coelenteramide.Synthesis and photolysis" Chemistry Lett.(1992)
[Publications] J.F.Rees,E.M.Thompson,F.Baquet and F.I.Tsuji: "Evidence for the utilization of coelenterazine as the luminescent substrate in Argyropelecus photophores" Molec.Bar.Biol.Biotech.1. 219-225 (1992)
[Publications] Y.Ohmiya,M.Ohashi and F.I.Tsuji: "Two excited states in scquorin bioluminescence induced by typrophan modification" FEBS Lett.301. 197-201 (1992)
[Publications] C.F.Qi,Y.Gomi,T.Hirano,M.Ohashi,Y.Ohmiya and F.I.Tsuji: "Chemi-and Bioluminescence of coclenterazine anlogues with phenyl homologues at the C-2position" J.Chem.Soc.Perkin Trans l. 1607-1611 (1992)
[Publications] S.Inouye and F.I.Tsuji: "Monitoring gene expression in Chinese hamster ovary cell using sccreted spoacquorin" Anal.Biochem.201. 114-118 (1992)
[Publications] I.Saito: "Photochecmistry of highly organized biomolecules:Scquence selective Photoreaction of DNA" Pure Appl.Chem.64. 1305-1310 (1992)
[Publications] H.Sugiyama,I.Saito et al: "Chemistry of neocarzinostatin-Mediated Cleavge of Oligonuclcotides" J.Am.Chem.Soc.114. 5573-5578 (1992)
[Publications] S.Kurono,T.Hirano,K.Tsujimoto,Y.Ohkawa,M.Yonede and M.Ohashi: "Delection of Molecular-ions of Cyelodextrin Inclusion Complexes" Org Mass Spectrom.27. 1157-1160 (1992)
[Publications] K.Tsujimoto,M.Aoki,M.Ohashi: "Stereoselective Photoisomerization of 4-retinal" J.Photochem.Phtobiol.A:Chem.65. 73-78 (1992)
[Publications] T.HIrano,Y.Gomi,T.Takahashi,K.Kitahara,F.Q.Chem.I.MIzoguchi,S.Kyushin,M.Ohashi: "Chemiluminescence of Coelenterazine Analogucs-Structures of Emitting Specics" Tetrahedron Letters. 33. 5771-5774 (1992)
[Publications] K.Teranishi,M.Isobe,T.Yamada and T.Goto: "Revision of Structure of Yellow Compound,a reduction Product from Acquorin,Photoprotein in Jellyfish,Aequorea acquorea" Tctrahcdron Letters. 33. 1303-1306 (1992)
[Publications] Y.Toya,M.Takagi,H.Nakata,N.suzuki,M.Isobe and T.Goto: "A Convenient Synthetic Method of 2-Cyano-l-methoxhbenzothiazole,A Key Intermediatc for the Synthesis of Firefly Luciferin" Bull.Chem.Soc.Japan. 65. 392-395 (1992)
[Publications] Y.Toya,M.Takagi,H.Nakata,N.Suzuki,M.Isobe and T.Goto: "Improved Synthetic Mcthods of Firefly Luciferin Derivatives for Use in Biojuminescent Anlysis of Hydrolytic Enzymes;Carboxylic Esterase and Alkaline Phosphatase" Bull.of Chem.Soc.Japan. 65. 2604-2610 (1992)
[Publications] I.Saito et al.: "Hydroxyl Radical Mediated Damage to Proteins,with special reference to the crystallins" Biochemistry. 31. 4296-4303 (1992)
[Publications] 磯部稔,近江谷克裕,F.I.辻,大橋守: "日本農芸化学会誌66No.4" 日本農芸化学会, 733-762 (1992)
[Publications] 編者水野卓,川合正允,著者磯部稔: "キノコの化学・生化学" 学会出版センター, 134-137 (1992)
[Publications] H.Nakayama,M.Taki,Y.Kanaoka: "Synthesis and propertics of diazipine,A novel photoaffinity probe of dihydropyridine receptors.Methods in Pharmacol.Vol.7(eds.H.Glossmann,J.Striessing)" Plenum Press,

1992 Fiscal Year Annual Research Report

励起分子の関与する精密構造認識と機能発現

Principal Investigator

大橋 守 電気通信大学, 電気通信学部, 教授 (70015535)

Research Products

[Publications] H.Nakayam,Y.Hatanaka,E.Yoshida,K.Oka,Y.Kanaoka et al.: "Photolabeled sites with a tetrodotosin dericative in the domain III and Iv of the electroplax sodium channel" Biochem.Biophys.Res.Commun.184. 900-907 (1992)

[Publications] Y.Hatanaka,H.Nakayama,Y.Kanaoka: "Photoaffinity labcling of the electroplax sodium channel with a photorcactive μ-conotoxin carring a radioactive and chromogenic derivative." Chem.Pharm.Bull.40. 2597-2539 (1992)

[Publications] A.Kuniyasu,K.Oka,T.Ide-Yamada,Y.,Hatanaka,H.Nakayama et al: "Structural characterization of the cardiac calcium channel from procine hearts" J.Biochem.112. 235-242 (1992)

[Publications] H.Nakayam,A.Yamamoto,Y.Hatanaka,T.Hachisu,S.Tsunasawa et al.: "Characterization of saccharide moiety in the electroplax sodium channel" Chem.Pharm.Bull.40. 3287-3291 (1992)

[Publications] H.Nakayama,H.Shikano,Y.Kanaoka: "A site directed antibody which inhibits phosphorylation of the rat brain sodium channel by CAMP-dependent protein kinase" Biochem.Biophys.Acta. 1175. 67-72 (1992)

[Publications] F.Q.Chen,T.Hirano,M.Ohashi,H.Nakayama,K.Oda,M.Machida: "3-Benzoy1-5-(4 hydroxyphenyl)-2-[4-(1-azi-2,2,2-trifluoroethyl)phenylacetanido]phrazine.A photosensitive analogue of coelenteramide.Synthesis and photolysis" Chemistry Lett.(1992)

[Publications] J.F.Rees,E.M.Thompson,F.Baquet and F.I.Tsuji: "Evidence for the utilization of coelenterazine as the luminescent substrate in Argyropelecus photophores" Molec.Bar.Biol.Biotech.1. 219-225 (1992)

[Publications] Y.Ohmiya,M.Ohashi and F.I.Tsuji: "Two excited states in scquorin bioluminescence induced by typrophan modification" FEBS Lett.301. 197-201 (1992)

[Publications] C.F.Qi,Y.Gomi,T.Hirano,M.Ohashi,Y.Ohmiya and F.I.Tsuji: "Chemi-and Bioluminescence of coclenterazine anlogues with phenyl homologues at the C-2position" J.Chem.Soc.Perkin Trans l. 1607-1611 (1992)

[Publications] S.Inouye and F.I.Tsuji: "Monitoring gene expression in Chinese hamster ovary cell using sccreted spoacquorin" Anal.Biochem.201. 114-118 (1992)

[Publications] I.Saito: "Photochecmistry of highly organized biomolecules:Scquence selective Photoreaction of DNA" Pure Appl.Chem.64. 1305-1310 (1992)

[Publications] H.Sugiyama,I.Saito et al: "Chemistry of neocarzinostatin-Mediated Cleavge of Oligonuclcotides" J.Am.Chem.Soc.114. 5573-5578 (1992)

[Publications] S.Kurono,T.Hirano,K.Tsujimoto,Y.Ohkawa,M.Yonede and M.Ohashi: "Delection of Molecular-ions of Cyelodextrin Inclusion Complexes" Org Mass Spectrom.27. 1157-1160 (1992)

[Publications] K.Tsujimoto,M.Aoki,M.Ohashi: "Stereoselective Photoisomerization of 4-retinal" J.Photochem.Phtobiol.A:Chem.65. 73-78 (1992)

[Publications] T.HIrano,Y.Gomi,T.Takahashi,K.Kitahara,F.Q.Chem.I.MIzoguchi,S.Kyushin,M.Ohashi: "Chemiluminescence of Coelenterazine Analogucs-Structures of Emitting Specics" Tetrahedron Letters. 33. 5771-5774 (1992)

[Publications] K.Teranishi,M.Isobe,T.Yamada and T.Goto: "Revision of Structure of Yellow Compound,a reduction Product from Acquorin,Photoprotein in Jellyfish,Aequorea acquorea" Tctrahcdron Letters. 33. 1303-1306 (1992)

[Publications] Y.Toya,M.Takagi,H.Nakata,N.suzuki,M.Isobe and T.Goto: "A Convenient Synthetic Method of 2-Cyano-l-methoxhbenzothiazole,A Key Intermediatc for the Synthesis of Firefly Luciferin" Bull.Chem.Soc.Japan. 65. 392-395 (1992)

[Publications] Y.Toya,M.Takagi,H.Nakata,N.Suzuki,M.Isobe and T.Goto: "Improved Synthetic Mcthods of Firefly Luciferin Derivatives for Use in Biojuminescent Anlysis of Hydrolytic Enzymes;Carboxylic Esterase and Alkaline Phosphatase" Bull.of Chem.Soc.Japan. 65. 2604-2610 (1992)

[Publications] I.Saito et al.: "Hydroxyl Radical Mediated Damage to Proteins,with special reference to the crystallins" Biochemistry. 31. 4296-4303 (1992)

[Publications] 磯部 稔,近江谷 克裕,F.I.辻,大橋 守: "日本農芸化学会誌66No.4" 日本農芸化学会, 733-762 (1992)

[Publications] 編者水野 卓,川合 正允,著者磯部 稔: "キノコの化学・生化学" 学会出版センター, 134-137 (1992)

[Publications] H.Nakayama,M.Taki,Y.Kanaoka: "Synthesis and propertics of diazipine,A novel photoaffinity probe of dihydropyridine receptors.Methods in Pharmacol.Vol.7(eds.H.Glossmann,J.Striessing)" Plenum Press,

大橋守電気通信大学, 電気通信学部, 教授 (70015535)

[Publications] 磯部稔,近江谷克裕,F.I.辻,大橋守: "日本農芸化学会誌66No.4" 日本農芸化学会, 733-762 (1992)

[Publications] 編者水野卓,川合正允,著者磯部稔: "キノコの化学・生化学" 学会出版センター, 134-137 (1992)