1996 年度実績報告書

液胞系オルガネラの多彩な機能と薬物の作用

研究課題

研究課題/領域番号	08307018
応募区分	総合
研究機関	大阪大学
研究代表者	二井將光大阪大学, 産業科学研究所, 教授 (50012646)
研究分担者	新井洋由東京大学, 薬学部, 助教授 (40167987) 森山芳則大阪大学, 産業科学研究所, 助教授 (10150658) 柿沼喜己千葉大学, 薬学部, 助教授 (80134394) 富岡清京都大学, 薬学部, 教授 (50114575) 前田正知大阪大学, 薬学部, 教授 (80190297)
キーワード	液胞 / 液胞ATPase / ベータ-ラクタム / bafilomycin / concanamycinA / 酸性オルガネラ / H^+-PPiase / ホスファチジルセリン
研究概要	動植物の細胞には、液胞(vacuoles)、エンドソーム、リソソーム、シナプス小胞、クロマフィン顆粒などのオルガネラが存在しており、多様な機能を担っている。いずれも内部は液胞型ATPaseが輸送するH^+によって酸性pHに保たれている。本研究では、異なる液胞系オルガネラを取り扱っている研究者が情報を交換し、技術を提供しあいながら共同研究を行ってきた。本年度に得られた主な成果を要約する。液胞に存在する新規なABC輸送蛋白のcDNAのクローニングに成功し、薬物輸送に果たす役割を明らかにしている。ベータ-ラクタム類の不斉触媒合成反応を開発し、新しいベータ-ラクタム抗生物質の液胞に対する効果を検討した。腸内連鎖球菌に見いだされたNa^+共役型の液胞ATPaseは、bafilomycin A1の阻害を受けないが、destruxin Bの作用を受けることを示した。膀胱表層細胞に存在する液胞型ATPaseが尿のHpを酸性に保っていることを示した。新規酸性オルガネラ(マイクロベジクル)の内分泌制御における意義を解明するため、マイクロベジクル上の神経伝達物質輸送系の性質を明らかにした。スカベンジャー受容体のひとつであるSRB-1が、肝臓、脂肪組織、副腎などに大量に発現していることを見出した。好熱菌のF_1-ATPaseのγサブユニットがATPの加水分解にともなって回転することを明らかにし、液胞型ATPaseにもあてはまることを推定した。さらにconcanamycinAによる阻害実験から、トランスゴルジネットワークの酸性化がglycosylphosphatidyl(GPI)-アンカーアルカリフォスファターゼのbasolateral側への輸送に重要であることが明らかになった。液胞膜輸送分子の解析を目的に、藻類シャジクモのH^+-PPiase、液胞型水チャネル2種、Ca^<2+>/H^+対抗輸送体のcDNA解析をし、液胞膜のCa^<2+>輸送機能を解析した。動物細胞の主要リン脂質の1つであるホスファチジルセリン(PS)の液胞系オルガネラの機能発現における役割を解析する目的で、PS合成酵素IIの欠損変異株を分離した。

研究成果
(48件)

すべてその他

すべて文献書誌 (48件)

[文献書誌] Y.Moriyama,et al.: "Role of endocrine cells microvesicles in intercellular chemical transduction." Biol.Chem.Hoppe-Seyler. 377. 155-165 (1996)
[文献書誌] K.Takeyasu,et al.: "Molecular imaging of Escherichia coli F_0F_1-ATPase in reconstituted membranes using atomic force microscopy." FEBS Lett.392. 110-113 (1996)
[文献書誌] H.Yamada,et al.: "Transmembrane topology of Escherichia coli H^+-ATPase(ATP synthase)subunit a." FEBS Lett.390. 34-38 (1996)
[文献書誌] M.Futai,et al.: "Conformational transmission in ATP synthase during catalysis:search for large structural changes." J.Bioenerg.and Biomemb.28(5). 409-414 (1996)
[文献書誌] K.Tomochika,et al.: "Vacuolar H^+-ATPase in mouse bladder epithelium is responsible for urinary acidification." FEBS Lett.404. 61-64 (1996)
[文献書誌] M.Muro,et al.: "Immunochemical identification of the pssA gene product as phosphatidylserine synthase I of Chinese hamster ovary cells." FEBS Letters. 395. 262-266 (1996)
[文献書誌] R.Sato,et al.: "Sterol-dependent transcriptional regulation of sterol regulatory element-binding protein-2." J.Biol.Chem.271. 26461-26464 (1996)
[文献書誌] M.Maeda,et al.: "Roles of gastric GATA DNA-binding proteins." J.Exp.Biol.199. 513-520 (1996)
[文献書誌] K.Tomioka,et al.: "Theoretical Calculation-Based Reproduction of Thermo-dynamic,Kinetic Behaviors and Cytotoxicity of Azasteganes." Heterocycles(Special Issue for the Memory of Prof.Yoshio Ban). 42(1). 43-45 (1996)
[文献書誌] M.Kanai,et al.: "Catalytic Asymmetric Conjugate Addition Reaction of Organocopper." 32(2). 171-174 (1996)
[文献書誌] M.Mizuno,et al.: "Chiral Ligand Controlled Enantioselective Opening of Oxirane and Oxetane" Tetrahedron:Asymmetry. 7(9). 2483-2484 (1996)
[文献書誌] Y.Nakagawa,et al.: "Structural Requirements of External,Chiral Amidophosphine Ligand for Asymmetric Reaction of Organocopper Reagent." Tetrahedron Letters. 37(43). 7805-7808 (1996)
[文献書誌] A.lida,et at.: "Trichothecinols A,B and C,Potent Anti-Tumor Promoting Sesquiterpenoids from the Fungus Trichothecium roseum." Tetrahedron Letters. 37(51). 9219-9220 (1996)
[文献書誌] T.Murata,et al.: "The ntpJ gene in the Enterococcus hirae ntp operon encodes a component of KtrII potassium transport system functionally independent of vacuolar Na^+-ATPase." J.Biol.Chem.271. 10042-10047 (1996)
[文献書誌] T.Murata,et al.: "Intracellular Na^+ reglulates transcription of the ntp operon encoding a vacuolar-type Na^+-translocating ATPase in Enterococcus hirae." J.Biol.Chem.271. 23661-23666 (1996)
[文献書誌] M.Hayashi,et al.: "Rapid increase in number of vanadocytes in the vanadium-rich ascidian,Ascidia sydneiensis samea upon treatment of live animals with ammonium chloride." J.Exp.Zool.27. 1-7 (1996)
[文献書誌] Y.Moriyama,et al.: "Vesicular monoamine transporter in microvesicles from bovine posterior pituitaries is immunologically similar to but distinct from the chromaffin granule counterpart in its sensitivities to 1-methyl-4-phenylpyridinium and histamine" Biochem.Biophys.Res.Commun.221. 790-794 (1996)
[文献書誌] H.Yamada,et al.: "Transmembrane topology of Escherichia coli H^+-ATPase(ATP synthase)subunit a." FEBS Letters. 390. 34-38 (1996)
[文献書誌] H.Yamada,et al.: "The L-type Ca^<2+> channel is involved in microvesicle-mediated glutamate exocytosis from pinealocytes." J.Pineal Research. 21. 165-174 (1996)
[文献書誌] H.Yamada,et al.: "Microvesicle-mediated exocytosis of glutamate is a novel paracrine-like chemical transduction mechanism and inhibits melatonin secretion in rat pinealocytes." J.Pineal Res.21. 175-191 (1996)
[文献書誌] I.Nakamura,et al.: "Failure of bone resorption in osteocleroic(oc/oc)mice is possibly due to the dissociation of vacuolar proton ATPase and cytoskeleton in osteoclasts." FEBS Letters. (in press). (1997)
[文献書誌] K.Tomochika,et al.: "Vacuolar proton ATPase in mouse bladder epithelium is responsible for urinary acidification." FEBS Letters. 404. 61-64 (1997)
[文献書誌] Y.Moriyama,et al.: "Membrane energization by proton pumps is inportant for compartmentation of drugs and toxins:a new concept for active transport." J.Exp.Biol.199. 1447-1454 (1996)
[文献書誌] Y.Moriyama,et al.: "Microvesicles as a cargo of novel intercellular chemical transduction from endocrine cells." Biol.Chem.Hoppe Sayler. 377. 155-165 (1996)
[文献書誌] Y.S.Ho,et al.: "Brain acetylhydrolase that inactivates platelet-activating factor is a G-protein-like trimer" Nature. 385. 89-93 (1997)
[文献書誌] T.Sato,et al.: "Serine phospholipid-specific phospholiopase A that is secreted from activated platelets" J.Biol.Chem.272. 2192-2198 (1997)
[文献書誌] K.Hattori,et al.: "cDNA cloning and expression of intracellular platelet-activating factor(PAF)acetylhydrolase II" J.Biol.Chem.271(51). 33032-33038 (1996)
[文献書誌] U.Albrecht,et al.: "Platelet-activating factor acetylhydrolase expression and activity suggest a link between neuroral migration and platelet-activating factor" Developmental Biology. 180. 579-593 (1996)
[文献書誌] M.Fukasawa,et al.: "SRB1,a class B scavenger receptor,recognizes both negatively charged liposomes and apoptotic cells." Exp.Cell Res.222. 246-250 (1996)
[文献書誌] T.Amano,et al.: "F_1-ATPase with 1,2,or3 incompetent catalytic sites." J.Biol.Chem.271. 17343-17348 (1996)
[文献書誌] K.Sato,et al.: "Characterization of the potential catalytic residue of the high affinity ATP-binding site of Escherichia coli SecA,translocation ATPase." J.Biol.Chem.271. 17439-17444 (1996)
[文献書誌] T.Matsui,et al.: "Catalytic activity of the α3β3γ complex of F_1-ATPase without noncatalytic nucleotide binding site." J.Biol.Chem.272(in press). (1997)
[文献書誌] H.Noji,et al.: "Direct observation of the rotation of F_1-ATPase." Nature. (in press). (1997)
[文献書誌] H.Itabe,et al.: "Sensitive detection of oxidatively modified low density lipoprotein using a monoclonal antibody." J.Lipid Res.37. 45-53 (1996)
[文献書誌] T.Imanaka,et al.: "Insertion of the 70-kDa peroxisomal membrane protein into peroxisomal membranes in vivo and in vitro." J.Biol.Chem.271. 3706-3713 (1996)
[文献書誌] Y.Takahashi,et al.: "Spatial and temporal pattern of smooth muscle cell differentiation during development of vascular system in the mouse embryo." Anat Embryol.194. 515-526 (1996)
[文献書誌] M.Mori,et al.: "Characterization of vitronectin in atherosclerotic lesions." J.Atheroscler.Thromb.3. 25-31 (1996)
[文献書誌] M.Arai,et al.: "Import into mitochondria of phospholipid hydroperoxide glutathione peroxidase requires a leader sequence." Biochem.Biophys.Res.Commun.227. 433-439 (1996)
[文献書誌] H.Itabe,et al.: "Oxidized phosphatidylcholines which modify proteins.Analysis by monoclonal antibody against oxidized low density lipoprotein." J.Biol.Chem.271. 33208-33217 (1996)
[文献書誌] D.G.Robinson,et al.: "Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons." Planta. 198. 95-103 (1996)
[文献書誌] H.Matsumoto,et al.: "Response of the plant root to aluminium stress:Analysis of the inhibition of the root elongation and changes in membrane function." J.Plant Res.109. 99-105 (1996)
[文献書誌] M.Maeshima,et al.: "Proton pumps of vacuolar membrane in growing plant cell." J.Plant Res.109. 119-125 (1996)
[文献書誌] T.Higuchi,et al.: "Complete nucleotide sequence of a cDNA for an intrinsic protein in vacuolar membranes from radish(Raphanus sativus)roots(PGR 96-045)." Plant Physiol.111. 947-947 (1996)
[文献書誌] T.Kitajima,et al.: "Novel bacterial rhodopsins from Haloarcula vallismortis." Biochem.Biophys.Res.Commun.220. 341-345 (1996)
[文献書誌] Y.Sugiyama,et al.: "Dual roles of DMPC and CHAPS in the refolding of bacterial opsins In Vitro." J.Biochem.119. 1143-1149 (1996)
[文献書誌] M.Shiozuka,et al.: "Stereo controlled synthesis of all stereoisomers of the proposed favolipin." Tetrahedron Letters. 37. 3875-3876 (1996)
[文献書誌] M,Shiozuka,et al.: "Synthesis of GLA-60 positional isomer as an LPS-agonist and its activity." Chemistry Letters. 735-736 (1996)
[文献書誌] K.Karasawa,et al.: "Cloning,expression and characterization of plasma platelet-activating factor-acetylhydrolase from the guinea pig." J.Biochem.120. 838-844 (1996)

1996 年度 実績報告書

液胞系オルガネラの多彩な機能と薬物の作用

研究代表者

二井 將光 大阪大学, 産業科学研究所, 教授 (50012646)

研究成果

[文献書誌] Y.Moriyama,et al.: "Role of endocrine cells microvesicles in intercellular chemical transduction." Biol.Chem.Hoppe-Seyler. 377. 155-165 (1996)

[文献書誌] K.Takeyasu,et al.: "Molecular imaging of Escherichia coli F_0F_1-ATPase in reconstituted membranes using atomic force microscopy." FEBS Lett.392. 110-113 (1996)

[文献書誌] H.Yamada,et al.: "Transmembrane topology of Escherichia coli H^+-ATPase(ATP synthase)subunit a." FEBS Lett.390. 34-38 (1996)

[文献書誌] M.Futai,et al.: "Conformational transmission in ATP synthase during catalysis:search for large structural changes." J.Bioenerg.and Biomemb.28(5). 409-414 (1996)

[文献書誌] K.Tomochika,et al.: "Vacuolar H^+-ATPase in mouse bladder epithelium is responsible for urinary acidification." FEBS Lett.404. 61-64 (1996)

[文献書誌] M.Muro,et al.: "Immunochemical identification of the pssA gene product as phosphatidylserine synthase I of Chinese hamster ovary cells." FEBS Letters. 395. 262-266 (1996)

[文献書誌] R.Sato,et al.: "Sterol-dependent transcriptional regulation of sterol regulatory element-binding protein-2." J.Biol.Chem.271. 26461-26464 (1996)

[文献書誌] M.Maeda,et al.: "Roles of gastric GATA DNA-binding proteins." J.Exp.Biol.199. 513-520 (1996)

[文献書誌] K.Tomioka,et al.: "Theoretical Calculation-Based Reproduction of Thermo-dynamic,Kinetic Behaviors and Cytotoxicity of Azasteganes." Heterocycles(Special Issue for the Memory of Prof.Yoshio Ban). 42(1). 43-45 (1996)

[文献書誌] M.Kanai,et al.: "Catalytic Asymmetric Conjugate Addition Reaction of Organocopper." 32(2). 171-174 (1996)

[文献書誌] M.Mizuno,et al.: "Chiral Ligand Controlled Enantioselective Opening of Oxirane and Oxetane" Tetrahedron:Asymmetry. 7(9). 2483-2484 (1996)

[文献書誌] Y.Nakagawa,et al.: "Structural Requirements of External,Chiral Amidophosphine Ligand for Asymmetric Reaction of Organocopper Reagent." Tetrahedron Letters. 37(43). 7805-7808 (1996)

[文献書誌] A.lida,et at.: "Trichothecinols A,B and C,Potent Anti-Tumor Promoting Sesquiterpenoids from the Fungus Trichothecium roseum." Tetrahedron Letters. 37(51). 9219-9220 (1996)

[文献書誌] T.Murata,et al.: "The ntpJ gene in the Enterococcus hirae ntp operon encodes a component of KtrII potassium transport system functionally independent of vacuolar Na^+-ATPase." J.Biol.Chem.271. 10042-10047 (1996)

[文献書誌] T.Murata,et al.: "Intracellular Na^+ reglulates transcription of the ntp operon encoding a vacuolar-type Na^+-translocating ATPase in Enterococcus hirae." J.Biol.Chem.271. 23661-23666 (1996)

[文献書誌] M.Hayashi,et al.: "Rapid increase in number of vanadocytes in the vanadium-rich ascidian,Ascidia sydneiensis samea upon treatment of live animals with ammonium chloride." J.Exp.Zool.27. 1-7 (1996)

[文献書誌] H.Yamada,et al.: "Transmembrane topology of Escherichia coli H^+-ATPase(ATP synthase)subunit a." FEBS Letters. 390. 34-38 (1996)

[文献書誌] H.Yamada,et al.: "The L-type Ca^<2+> channel is involved in microvesicle-mediated glutamate exocytosis from pinealocytes." J.Pineal Research. 21. 165-174 (1996)

[文献書誌] H.Yamada,et al.: "Microvesicle-mediated exocytosis of glutamate is a novel paracrine-like chemical transduction mechanism and inhibits melatonin secretion in rat pinealocytes." J.Pineal Res.21. 175-191 (1996)

[文献書誌] I.Nakamura,et al.: "Failure of bone resorption in osteocleroic(oc/oc)mice is possibly due to the dissociation of vacuolar proton ATPase and cytoskeleton in osteoclasts." FEBS Letters. (in press). (1997)

[文献書誌] K.Tomochika,et al.: "Vacuolar proton ATPase in mouse bladder epithelium is responsible for urinary acidification." FEBS Letters. 404. 61-64 (1997)

[文献書誌] Y.Moriyama,et al.: "Membrane energization by proton pumps is inportant for compartmentation of drugs and toxins:a new concept for active transport." J.Exp.Biol.199. 1447-1454 (1996)

[文献書誌] Y.Moriyama,et al.: "Microvesicles as a cargo of novel intercellular chemical transduction from endocrine cells." Biol.Chem.Hoppe Sayler. 377. 155-165 (1996)

[文献書誌] Y.S.Ho,et al.: "Brain acetylhydrolase that inactivates platelet-activating factor is a G-protein-like trimer" Nature. 385. 89-93 (1997)

[文献書誌] T.Sato,et al.: "Serine phospholipid-specific phospholiopase A that is secreted from activated platelets" J.Biol.Chem.272. 2192-2198 (1997)

[文献書誌] K.Hattori,et al.: "cDNA cloning and expression of intracellular platelet-activating factor(PAF)acetylhydrolase II" J.Biol.Chem.271(51). 33032-33038 (1996)

[文献書誌] U.Albrecht,et al.: "Platelet-activating factor acetylhydrolase expression and activity suggest a link between neuroral migration and platelet-activating factor" Developmental Biology. 180. 579-593 (1996)

[文献書誌] M.Fukasawa,et al.: "SRB1,a class B scavenger receptor,recognizes both negatively charged liposomes and apoptotic cells." Exp.Cell Res.222. 246-250 (1996)

[文献書誌] T.Amano,et al.: "F_1-ATPase with 1,2,or3 incompetent catalytic sites." J.Biol.Chem.271. 17343-17348 (1996)

[文献書誌] K.Sato,et al.: "Characterization of the potential catalytic residue of the high affinity ATP-binding site of Escherichia coli SecA,translocation ATPase." J.Biol.Chem.271. 17439-17444 (1996)

[文献書誌] T.Matsui,et al.: "Catalytic activity of the α3β3γ complex of F_1-ATPase without noncatalytic nucleotide binding site." J.Biol.Chem.272(in press). (1997)

[文献書誌] H.Noji,et al.: "Direct observation of the rotation of F_1-ATPase." Nature. (in press). (1997)

[文献書誌] H.Itabe,et al.: "Sensitive detection of oxidatively modified low density lipoprotein using a monoclonal antibody." J.Lipid Res.37. 45-53 (1996)

[文献書誌] T.Imanaka,et al.: "Insertion of the 70-kDa peroxisomal membrane protein into peroxisomal membranes in vivo and in vitro." J.Biol.Chem.271. 3706-3713 (1996)

[文献書誌] Y.Takahashi,et al.: "Spatial and temporal pattern of smooth muscle cell differentiation during development of vascular system in the mouse embryo." Anat Embryol.194. 515-526 (1996)

[文献書誌] M.Mori,et al.: "Characterization of vitronectin in atherosclerotic lesions." J.Atheroscler.Thromb.3. 25-31 (1996)

[文献書誌] M.Arai,et al.: "Import into mitochondria of phospholipid hydroperoxide glutathione peroxidase requires a leader sequence." Biochem.Biophys.Res.Commun.227. 433-439 (1996)

[文献書誌] H.Itabe,et al.: "Oxidized phosphatidylcholines which modify proteins.Analysis by monoclonal antibody against oxidized low density lipoprotein." J.Biol.Chem.271. 33208-33217 (1996)

[文献書誌] D.G.Robinson,et al.: "Immunological detection of tonoplast polypeptides in the plasma membrane of pea cotyledons." Planta. 198. 95-103 (1996)

[文献書誌] H.Matsumoto,et al.: "Response of the plant root to aluminium stress:Analysis of the inhibition of the root elongation and changes in membrane function." J.Plant Res.109. 99-105 (1996)

[文献書誌] M.Maeshima,et al.: "Proton pumps of vacuolar membrane in growing plant cell." J.Plant Res.109. 119-125 (1996)

[文献書誌] T.Higuchi,et al.: "Complete nucleotide sequence of a cDNA for an intrinsic protein in vacuolar membranes from radish(Raphanus sativus)roots(PGR 96-045)." Plant Physiol.111. 947-947 (1996)

[文献書誌] T.Kitajima,et al.: "Novel bacterial rhodopsins from Haloarcula vallismortis." Biochem.Biophys.Res.Commun.220. 341-345 (1996)

[文献書誌] Y.Sugiyama,et al.: "Dual roles of DMPC and CHAPS in the refolding of bacterial opsins In Vitro." J.Biochem.119. 1143-1149 (1996)

[文献書誌] M.Shiozuka,et al.: "Stereo controlled synthesis of all stereoisomers of the proposed favolipin." Tetrahedron Letters. 37. 3875-3876 (1996)

[文献書誌] M,Shiozuka,et al.: "Synthesis of GLA-60 positional isomer as an LPS-agonist and its activity." Chemistry Letters. 735-736 (1996)

[文献書誌] K.Karasawa,et al.: "Cloning,expression and characterization of plasma platelet-activating factor-acetylhydrolase from the guinea pig." J.Biochem.120. 838-844 (1996)

1996 年度実績報告書

二井將光大阪大学, 産業科学研究所, 教授 (50012646)