1992 Fiscal Year Annual Research Report

ATP合成酵素の高次構造

Research Project

Project/Area Number	04266104
Research Institution	Osaka University
Principal Investigator	二井將光大阪大学, 産業科学研究所, 教授 (50012646)
Co-Investigator(Kenkyū-buntansha)	茂木立志東京大学, 理学部, 助手 (90219965) 太田成男自治医科大学, 医学部, 助教授 (00125832) 白木原康雄兵庫教育大学, 学校教育学部, 助教授 (20150287) 伊藤雄而東大物性研, 中性子回折物性部門, 助教授 (10013517) 小澤高将名古屋大学, 医学部, 教授 (80022771) 田川邦夫大阪大学, 医学部, 教授 (40028296)
Keywords	ATP合成酵素 / 活性中心 / H^+輸送路 / αβ複合体
Research Abstract	ATPは生物においてエネルギー通貨としての役割を果たしている。このATPを合成しているATP合成酵素は生物に普遍的に存在している。基本的には本酵素はα,β,γ,δ,ε,a,b,cの8種のサブユニットからなることが確定している。本研究はATP合成酵素の高次構造を明らかにし、これに基づいて反応機構を解明することを究極の目的としている。今年度は本研究の初年度として、多くの基礎的な知見を得た。以下に得られた成果をまとめる。(1)ATP合成酵素のβサブユニットの活性中心の動的構造、特にATP結合部位のγ位リン酸基近傍の構造を確定した。さらにγサブユニットの機能部位、特にMet-23残基近傍とカルボキシル末端側(Gln-269→Thr-273)がH^+輸送とATP合成の共役に関与していることを明らかにした(二井)。(2)ミトコンドリアのATP合成酵素制御タンパクの結合部位を明らかにした(田川)。(3)H^+輸送路を形成するaサブユニットの疾病による構造変化を明らかにした(小澤)。この知見をふまえ大腸菌aサブユニットの構造を解析する基礎を作った(二井)。(4)大腸菌のATP合成酵素の構造を中性子散乱により明らかにするための基本的な条件を検討した(伊藤)。なお二井はこの研究のため大腸菌ATP合成酵素を供給した。(5)好熱菌のATP合成酵素のαβ複合体の結晶化に成功した(白木原)。(6)好熱菌のα3β3およびαβ複合体の動的変換の機構を解析し、X線による構造解析のための条件を決定した(太田)。ミトコンドリアのβサブユニット遺伝子の発現機構を解析した(太田)。(7)チトクロームとATP合成酵素のH^+輸送機構を明らかにする基礎を作った(茂木)。以上のように次年度の飛躍に向け、充分な成果が得られ、共同研究の準備も完成した。

Research Products
(39 results)

All Other

All Publications (39 results)

[Publications] A.Iwamoto: "Domains near ATP phosphate in the catalytic site of H^+-ATPase:Model proposed from mutagenesis and inhibitor studies" J.Biol.Chem.286. (1993)
[Publications] R.K.Nakamoto: "The γ subunit of the Escherichia coli ATP synthase:Mutations in the carboxyl-terminal region restore energy coupling to the amino-terminal mutant γ Met-23→Lys" J.Biol.Chem.286. 867-872 (1993)
[Publications] K.Shin: "F_0F_1-ATPase β subunit mutations perturb the coupling between catalysis and transport" J.Biol.Chem.267. 20835-20839 (1992)
[Publications] R.K.Nakamoto: "Escherichia coli F_0F_1-ATPase:residues inbolved in catalysis and coupling" Anal.N.Y.Acad.Sci.671. 335-344 (1992)
[Publications] H.Omote: "Effects mutations of conserved Lys-155 and Thr-156 residues in the phosphate binding glycine-rich sequence of the F_1-ATPase β subunit of Escherichia coli" J.Biol.Chem.267. 20571-20576 (1992)
[Publications] M.Futai: "Escherichia coli ATP sythase(F-Type ATPase):Catalytic site and regulation of H^+ translocation." J.Exp.Biol.172. 443-449 (1992)
[Publications] M.Futai: "A glycine-rich sequence in the catalytic site of F-type ATPase." J.Biomemb Bioenerg.24. 463-467 (1992)
[Publications] M.Futai: "Catalytic site in the β subunit of H^+-ATPase from Escherichia coli:Glycine-rich sequence and its interaction with Ser-174." Acta Physiol Scan.146. 125-130 (1992)
[Publications] M.Jounouchi: "Role of the amino terminal region of the δ subunit of Escherichia coli H^+-ATPase(F_0F_1)." Arch.Biochem.Biophys.292. 87-94 (1992)
[Publications] M.Jounouchi: "Escherichia coli H^+-ATPase:Role of the ε subunit in binding F_1 to the F_0 sector." Arch.Biochem.Biophys.292. 376-381 (1992)
[Publications] H.Mimura: "Binding of an intrinsic ATPase inhibitor to the interface between α-and β-subunits of F_0F_1 ATPase upon de-energization of mitochondria" J.Biochem.113. (1993)
[Publications] K.Torii: "Age-related decrease in repiratory muscle mitochondrial function in rats" Am.J.Respir.Cell Mol.Biol.6. 88-92 (1992)
[Publications] H.Yamamoto: "Signiflcant existence of deleted mitochondrial DNA in cirrhotic liver" Biochem.Biophys.Res.Commun.182. 88-92 (1992)
[Publications] K.Torii: "Aging-associated deletions of humman diapharagmatic mitochondrial DNA" Am.J.Respir.Cell Mol.Biol.6. 543-549 (1992)
[Publications] T.Ito: "Mitochondrial DNA mutations in cardiomyopathy" Jpn.Circ.J.56. 1045-1053 (1992)
[Publications] K.Sahashi: "Increased mitochondrial DNA deletions in the skeltal muscle of myotonic dystrophy" Gerontology. 38. 18-29 (1992)
[Publications] T.Obayashi: "Point mutations in mitochondrial DNA in patients with hypertrophic cardiomyopathy" Am.Heart J.24. 1263-1269 (1992)
[Publications] M.Kanai: "Mitochondrial dysfunction in the non-obstructed lobe of rat liver after selective bailary obstruction" Hepato-Gastroenterrology. 39. 385-391 (1992)
[Publications] M.Yahakawa: "Age-associated oxygen damage and mutations in mithocondrial DNA in human hearts" Biochem Biophys Res Commun.189. 979-985 (1992)
[Publications] Y.Ito: "Application of neutrons(small-angle scattering)" Radioisotopes. 41. 514-524 (1992)
[Publications] Y.Kagawa: "The αβ complexes of ATP synthase:the α_3β_3 oligomer and α1β1 protomer" J.Bioenergetics and Biomembranes. 24. 441-445 (1992)
[Publications] Y.Ito: "Comparison of small-angle neutron and X-ray scattering from insect lipophorin" Abstracts of the 1st neutron symposium Tokai,Japan 1992. 21 (1992)
[Publications] Y.Kagawa: "The α3β3 and α1β1 complexes of ATP synthase" Ann.New York Acad.Sci.671. 366-376 (1992)
[Publications] Y.Kodayashi: "The mutant mitochondnal genes in mitochondrial myopathy encephalopathy,lactic acidosis and stroke-like episodes(MELAS)were selectively amplified through generations." J.Inher.Metab.Dis.15. 803-808 (1992)
[Publications] K.Tominaga: "Upstream region of a genomic gene for human mitochondrial transcription factor 1" Biochim.Biophys.Acta. 1131. 217-219 (1992)
[Publications] S.Matsuda: "Molecular cloning of dihydrolipoamide acetyltransferase of the rat pyruvate dehydrogenase complex:sequence dihydrollpoamide acetyltrana-ferase" Biochim.Biophys.Acta. 1131. 114-118 (1992)
[Publications] Y.Kagawa: "Mitochondirial myopathy and transcriptional control" Med.Sports Sci.37. 324-328 (1992)
[Publications] J.Minagawa: "Identification of heme and copper ligands in subunit I of the cytochrome bo complex in Escherichia coli." J.Biol.Chem.286. (1993)
[Publications] K.Saiki: "Heme O beoshythesis in Escherichia coli;the cyoE gene in the cytochrome bo operon encodes a protoheme IX farnesyltransferase" Biochim.Biophys.Res.Commun.189. (1993)
[Publications] M.Tsubaki: "Structure of heme-copper binuclear center of the cytochrome bo complex in Escherichia coli;EPR and Fourier-transform infrared spectroscopic studies" Biochemistry in press. 32. (1993)
[Publications] 二井将光: "「生物はどのようにして物質を合成するのか」基礎生物学構座2 生体の物質的成り立ちと代謝" 朝倉書店, 55 (1993)
[Publications] 二井将光: "「部位指定突然変異」生命工学シリーズIV" 裳華房, 36 (1992)
[Publications] 二井将光: "「H^+-ATPase (F_0F_1-ATPase)」新生化学実験講座1 タンパク質VII タンパク質工学" 東京化学同人, 36 (1992)
[Publications] T.Ozawa: "Metabolic and Molecular Aspects of Cardiomyopathy" University of Tokyo Press, 20 (1992)
[Publications] T.Ozawa: "Molecular Biology of the Myocardium" Japan Scientific Press&CRC Press, 14 (1992)
[Publications] T.Ozawa: "New Aspects in the Treatment of Failing Heart" Springer-Verlag, 29 (1992)
[Publications] T.Ozawa: "Techniques in Molecular Neurobiology" Human Press, 29 (1992)
[Publications] 太田成男: "「ミトコンドリア遺伝子と核遺伝子の関係」代謝29" 17 (1992)
[Publications] 太田成男: "「ミトコンドリア遺伝子の点突然変異による疾患」細胞工学11" 7 (1992)

1992 Fiscal Year Annual Research Report

ATP合成酵素の高次構造

Principal Investigator

二井 將光 大阪大学, 産業科学研究所, 教授 (50012646)

Research Products

[Publications] A.Iwamoto: "Domains near ATP phosphate in the catalytic site of H^+-ATPase:Model proposed from mutagenesis and inhibitor studies" J.Biol.Chem.286. (1993)

[Publications] R.K.Nakamoto: "The γ subunit of the Escherichia coli ATP synthase:Mutations in the carboxyl-terminal region restore energy coupling to the amino-terminal mutant γ Met-23→Lys" J.Biol.Chem.286. 867-872 (1993)

[Publications] K.Shin: "F_0F_1-ATPase β subunit mutations perturb the coupling between catalysis and transport" J.Biol.Chem.267. 20835-20839 (1992)

[Publications] R.K.Nakamoto: "Escherichia coli F_0F_1-ATPase:residues inbolved in catalysis and coupling" Anal.N.Y.Acad.Sci.671. 335-344 (1992)

[Publications] H.Omote: "Effects mutations of conserved Lys-155 and Thr-156 residues in the phosphate binding glycine-rich sequence of the F_1-ATPase β subunit of Escherichia coli" J.Biol.Chem.267. 20571-20576 (1992)

[Publications] M.Futai: "Escherichia coli ATP sythase(F-Type ATPase):Catalytic site and regulation of H^+ translocation." J.Exp.Biol.172. 443-449 (1992)

[Publications] M.Futai: "A glycine-rich sequence in the catalytic site of F-type ATPase." J.Biomemb Bioenerg.24. 463-467 (1992)

[Publications] M.Futai: "Catalytic site in the β subunit of H^+-ATPase from Escherichia coli:Glycine-rich sequence and its interaction with Ser-174." Acta Physiol Scan.146. 125-130 (1992)

[Publications] M.Jounouchi: "Role of the amino terminal region of the δ subunit of Escherichia coli H^+-ATPase(F_0F_1)." Arch.Biochem.Biophys.292. 87-94 (1992)

[Publications] M.Jounouchi: "Escherichia coli H^+-ATPase:Role of the ε subunit in binding F_1 to the F_0 sector." Arch.Biochem.Biophys.292. 376-381 (1992)

[Publications] H.Mimura: "Binding of an intrinsic ATPase inhibitor to the interface between α-and β-subunits of F_0F_1 ATPase upon de-energization of mitochondria" J.Biochem.113. (1993)

[Publications] K.Torii: "Age-related decrease in repiratory muscle mitochondrial function in rats" Am.J.Respir.Cell Mol.Biol.6. 88-92 (1992)

[Publications] H.Yamamoto: "Signiflcant existence of deleted mitochondrial DNA in cirrhotic liver" Biochem.Biophys.Res.Commun.182. 88-92 (1992)

[Publications] K.Torii: "Aging-associated deletions of humman diapharagmatic mitochondrial DNA" Am.J.Respir.Cell Mol.Biol.6. 543-549 (1992)

[Publications] T.Ito: "Mitochondrial DNA mutations in cardiomyopathy" Jpn.Circ.J.56. 1045-1053 (1992)

[Publications] K.Sahashi: "Increased mitochondrial DNA deletions in the skeltal muscle of myotonic dystrophy" Gerontology. 38. 18-29 (1992)

[Publications] T.Obayashi: "Point mutations in mitochondrial DNA in patients with hypertrophic cardiomyopathy" Am.Heart J.24. 1263-1269 (1992)

[Publications] M.Kanai: "Mitochondrial dysfunction in the non-obstructed lobe of rat liver after selective bailary obstruction" Hepato-Gastroenterrology. 39. 385-391 (1992)

[Publications] M.Yahakawa: "Age-associated oxygen damage and mutations in mithocondrial DNA in human hearts" Biochem Biophys Res Commun.189. 979-985 (1992)

[Publications] Y.Ito: "Application of neutrons(small-angle scattering)" Radioisotopes. 41. 514-524 (1992)

[Publications] Y.Kagawa: "The αβ complexes of ATP synthase:the α_3β_3 oligomer and α1β1 protomer" J.Bioenergetics and Biomembranes. 24. 441-445 (1992)

[Publications] Y.Ito: "Comparison of small-angle neutron and X-ray scattering from insect lipophorin" Abstracts of the 1st neutron symposium Tokai,Japan 1992. 21 (1992)

[Publications] Y.Kagawa: "The α3β3 and α1β1 complexes of ATP synthase" Ann.New York Acad.Sci.671. 366-376 (1992)

[Publications] Y.Kodayashi: "The mutant mitochondnal genes in mitochondrial myopathy encephalopathy,lactic acidosis and stroke-like episodes(MELAS)were selectively amplified through generations." J.Inher.Metab.Dis.15. 803-808 (1992)

[Publications] K.Tominaga: "Upstream region of a genomic gene for human mitochondrial transcription factor 1" Biochim.Biophys.Acta. 1131. 217-219 (1992)

[Publications] S.Matsuda: "Molecular cloning of dihydrolipoamide acetyltransferase of the rat pyruvate dehydrogenase complex:sequence dihydrollpoamide acetyltrana-ferase" Biochim.Biophys.Acta. 1131. 114-118 (1992)

[Publications] Y.Kagawa: "Mitochondirial myopathy and transcriptional control" Med.Sports Sci.37. 324-328 (1992)

[Publications] J.Minagawa: "Identification of heme and copper ligands in subunit I of the cytochrome bo complex in Escherichia coli." J.Biol.Chem.286. (1993)

[Publications] K.Saiki: "Heme O beoshythesis in Escherichia coli;the cyoE gene in the cytochrome bo operon encodes a protoheme IX farnesyltransferase" Biochim.Biophys.Res.Commun.189. (1993)

[Publications] M.Tsubaki: "Structure of heme-copper binuclear center of the cytochrome bo complex in Escherichia coli;EPR and Fourier-transform infrared spectroscopic studies" Biochemistry in press. 32. (1993)

[Publications] 二井 将光: "「生物はどのようにして物質を合成するのか」基礎生物学構座2 生体の物質的成り立ちと代謝" 朝倉書店, 55 (1993)

[Publications] 二井 将光: "「部位指定突然変異」生命工学シリーズIV" 裳華房, 36 (1992)

[Publications] 二井 将光: "「H^+-ATPase (F_0F_1-ATPase)」 新生化学実験講座1 タンパク質VII タンパク質工学" 東京化学同人, 36 (1992)

[Publications] T.Ozawa: "Metabolic and Molecular Aspects of Cardiomyopathy" University of Tokyo Press, 20 (1992)

[Publications] T.Ozawa: "Molecular Biology of the Myocardium" Japan Scientific Press&CRC Press, 14 (1992)

[Publications] T.Ozawa: "New Aspects in the Treatment of Failing Heart" Springer-Verlag, 29 (1992)

[Publications] T.Ozawa: "Techniques in Molecular Neurobiology" Human Press, 29 (1992)

[Publications] 太田 成男: "「ミトコンドリア遺伝子と核遺伝子の関係」代謝29" 17 (1992)

[Publications] 太田 成男: "「ミトコンドリア遺伝子の点突然変異による疾患」細胞工学11" 7 (1992)

二井將光大阪大学, 産業科学研究所, 教授 (50012646)

[Publications] 二井将光: "「生物はどのようにして物質を合成するのか」基礎生物学構座2 生体の物質的成り立ちと代謝" 朝倉書店, 55 (1993)

[Publications] 二井将光: "「部位指定突然変異」生命工学シリーズIV" 裳華房, 36 (1992)

[Publications] 二井将光: "「H^+-ATPase (F_0F_1-ATPase)」新生化学実験講座1 タンパク質VII タンパク質工学" 東京化学同人, 36 (1992)

[Publications] 太田成男: "「ミトコンドリア遺伝子と核遺伝子の関係」代謝29" 17 (1992)

[Publications] 太田成男: "「ミトコンドリア遺伝子の点突然変異による疾患」細胞工学11" 7 (1992)