細胞エネルギー共役系の構築と調節

1993 Fiscal Year Annual Research Report

• Project/Area Number: 04266102
• Research Institution: The University of Tokyo
• Principal Investigator: 安楽 泰宏 東京大学, 大学院・理学系研究科, 教授 (20012643)
• Co-Investigator(Kenkyū-buntansha): 向畑 泰男 名古屋大学, 理学部, 教授 (10028110) ; 川喜田 正夫 東京都臨床医学総合研究所, 医化学研究部, 部長 (00012740) ; 松原 央 大阪大学, 理学部, 教授 (00028242) ; 吉田 賢右 東京工業大学, 資源化学研究所, 教授 (90049073) ; 二井 將光 大阪大学, 産業科学研究所, 教授 (50012646)
• Keywords: 細胞エネルギー / ATP合成酵素 / 液胞型ATP分解酵素 / 呼吸酵素 / 電子伝達複合体 / 輸送蛋白 / オスモエンザイム / 化学浸透学説

Research Abstract

本総括班は、重点領域研究「細胞エネルギー」の研究目的を遅滞無く達成させるため、計画研究の推進、班間・班員間の研究連絡と調整、研究成果の評価などの重要業務をおこなった。
1.本年度は2件の公開講演会を行った。1)日本生化学会シンポジウム[細胞の機能とオスモエンザイム](平成5年10月4日:ATPaseに関する演題6題)。2)公開講演会[呼吸鎖電子伝達系の構造と機能](平成5年12月13日:呼吸鎖電子伝達系末端酸化酵素の構造化学、反応中心、電子伝達機構に関する演題10題)。両シンポジウムとも多数の聴衆をあつめ盛会であった。
2.合同班会議(平成5年12月14-16日:大阪千里ライフサイエンスセンター)を開催した、全班員の研究業績を聴取し、討論を行った。
3.総括班会議(平成5年12月13日)を開催し、平成5年度の研究推進の方針およびその経過について報告した。これに基ずいて平成6年度の研究・運営方針につき討議をおこなった。総括班員は合同班会議に出席し、研究業績の評価・指導をおこなった。
4.平成5年度の全成果を取りまとめ、[細胞エネルギー共役系の構築と調節]の研究成果報告書を出版した。ニュースレターを3回発行し、班員に配布した。特に、国際会議に出席し講演を行った班員の研究成果を班員に周知した。
5.Paul D.Bover博士特別講演会を開催し〔実行委員長 吉田賢右教授:平成5年4月21日)、ATP合成酵素の作動原理に関する討論を行い、意見を交換した。

Research Products

• [Publications] Y.Anraku: "Protoenzyme:Emerging evidence in nature." J.Biochem.115. 175-178 (1994)
• [Publications] M.N.Ho: "VMA13 encodes a 54-kDa vacuolar H^+-ATPase subunit required for activity but not assembly of the enzyme complex in Saccharomyces cerevisiae." J.Biol.Chem.268. 18286-18292 (1993)
• [Publications] R.Hirata: "VMA12 is essential for assembly of the vacuolar H^+-ATPase subunits onto the vacuolar membrane in Saccharomyces cerevisiae." J.Biol.Chem.268. 961-967 (1993)
• [Publications] N.Ishii: "Three dimensional structure of F_1-ATPase of thermophilic bacterium PS3 obtained from electron crystallography." J.Biochem.113. 245-250 (1993)
• [Publications] C.Kaibara: "AT(D)P Mg-induced dissociation of the α_3β_3 complex of the F_1-ATPase from a thermophilic Bacillus PS3 into α_1β_1 heterodimers is prevented by mutation β(Y341C)." FEBS Lett.321. 46-50 (1993)
• [Publications] K.Tozawa: "Structure of the α subunit of F_1-ATPase probed by limited proteolysis." J.Biol.Chem.268. 19044-19054 (1993)
• [Publications] E.Muneyuki: "Inhibitory effect of NaN_3 on the F_0F_1 ATPase of submitochondrial particles as related to nucleotide binding." Biochim.Biophys.Acta. 1144. 62-68 (1993)
• [Publications] E.Muneyuki: "Direct transmembraneous reconstitution of bacteriorhodopsin into planar phospholipid bilayers." Biochim.Biophys.Acta. 1183. 171-179 (1993)
• [Publications] M.Odaka: "In vitro affinity label of a protein expressed in Escherichia coli;Coenzyme A occupied the AT(D)P binding site of the mutant F_1-ATPase β subunit(Y307C)through a disulfide bond." FEBS Lett.336. 231-235 (1993)
• [Publications] A.Iwamoto: "Domains near ATP γ phosphate in the catalytic site of H^+-ATPase:Model proposed from mutagenesis and inhibitor studies." J.Biol.Chem.268. 3156-3160 (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.268. 867-872 (1993)
• [Publications] D.Xie: "The atp2 operon of the green bacterium Chlorobium limicola." Biochim.Biophys.Acta. 1172. 267-273 (1993)
• [Publications] M.Nakai: "Acidic regions of cytochrome c_1 are essential for ubiquinol-cytochrome c reductase activity in yeast cells lacking the acidic QCR6 protein." J.Biochem.114. 919-925 (1993)
• [Publications] M.Nakai: "Intramitochondrial protein sorting:Isolation and characterization of the yeast MSP1 gene which belongs to a novel family of putative ATPases." J.Biol.Chem.268. 24262-24269 (1993)
• [Publications] J.-Y.Cui: "Molecular cloning and nucleotide sequences of cDNAs encoding subunits I,II,and IX of Euglena gracilis mitochondrial complex III." J.Biochem.115. 98-107 (1994)
• [Publications] H.Oh-oka: "Isolation of the photoactive reaction center complex that contains three types of Fe-S centers and a cytochrome c subunit from the green sulfur bacterium Chlorobium limicola f.thiosulfatophilum,strain Larsen." Plant Cell Physiol.34. 93-101 (1993)
• [Publications] N.Kunishima: "Crystallization and preliminary X-ray diffraction studies of peroxidase from a fungus Arthromyces ramosus." Proteins,Structures,Functions,and Genetics. 15. 216-220 (1993)
• [Publications] K.Saeki: "Nucleotide sequence and genetic analysis of the region essential for functional expression of the gene for ferredoxin I,fdxN,in Rhodobacter capsulatus:Sharing of one upstream activator sequence in opposite directions by two operons related to nitrogen fixation." Plant Cell Physiol.34. 185-199 (1993)
• [Publications] Y.Fujita: "Identification of a nifDK-like gene(ORF467)involved in the biosynthesis of chlorophyll in the cyanobacterium Plectonema boryanum." Plant Cell Physiol.34. 305-314 (1993)
• [Publications] T.Yamashita: "Characterization of the Na^+/H^+-antiporter of bovine renal cortex and its immunoaffinity purification in a reconstitutively active form." J.Biochem.113. 136-140 (1993)
• [Publications] T.Suzuki: "Uncoupling of ATP splitting from Ca^<2+>-transport in Ca^<2+>-transporting ATPase of the sarcoplasmic reticulum as a result of modification by N-(3-pyrene)maleimide:Activation of a channel with a specificity for alkaline earth metal ions." J.Biochem.114. 203-209 (1993)
• [Publications] M.Kuroda: "Calcium pump expression in human bone and soft tissue tumors." Virchows Archiv.A Pathol.Anat.423. 209-213 (1993)
• [Publications] T.Hara: "The UDP-galactose translocator gene is mapped to the Wiskott-Aldrich syndrome locus of the X chromosome." Somat.Cell Mol.Genet.(in press).
• [Publications] C.Matsuda: "Gene structure of human mitochondrial ATP synthase γ-subunit:Tissue-specificity produced by alternative RNA splicing." J.Biol.Chem.268. 24950-24958 (1993)
• [Publications] Y.Takiyama: "The gene for Machado-Joseph disease maps to human chromosome 14q." Nature Genetics. 4. 300-304 (1993)
• [Publications] T.Ohta: "Heat shock promoter of thermophilic chaperonin operon." Biochem.Biophys.Res.Commun.191. 550-557 (1993)
• [Publications] Y.Sunada: "Inherited amyloid polyneuropathy type IV(gelsolin variant)in a Japanese family." Ann.Neurol.33. 57-62 (1993)
• [Publications] M.Nakanishi: "Solubilization and reconstitution of high-and low-affinity Na^+-dependent neutral L-α-amino acid transporters from rabbit small intestine." Biochim.Biophys.Acta. 1151. 193-200 (1993)
• [Publications] C.Matsuda: "Tissue-specific isoforms of the bovine mitochondrial ATP synthase γ-subunit." FEBS Lett.325. 281-284 (1993)
• [Publications] H.-J.Schaefer: "Photocrosslinking of ATP synthase." Biol.Chem.Hoppe-Seyler. 674. 164 (1993)
• [Publications] M.Yuzaki: "Cerebellar glia specifically supports the survival of Purkinje cells in culture." Biochem.Biophys.Res.Commun.197. 123-129 (1993)
• [Publications] K.Tominaga: "Smaller isoform of human mitochondrial transcription factor 1:Its wide distribution and production by alternative splicing." Biochem.Biophys.Res.Commun.194. 544-551 (1993)
• [Publications] A.Schuhen: "Fluorescent bifunctional nucleotide analogs." Biol.Chem.Hoppe-Seyler. 674. 164 (1993)
• [Publications] N.Shiraiwa: "Content of mutant mitochondrial DNA and organ-dysfunction in a patient with a MELAS-subgroup of mitochondrial encephalomyopathies." J.Neurol.Sci.in press.
• [Publications] M.Nakanishi: "Purification and reconstitution of an intestinal Na^+-dependent neutral L-α-amino acid transporter." J.Biol.Chem.269(in press). (1994)
• [Publications] J-I.Hayashi: "Nuclear but not mitochondrial genome involvement in human age-relateed mitochondrial dysfunction:Funcional integrity of mitochondrial DNA from aged subjects." J.Biol.Chem.269(in press). (1994)
• [Publications] S.S.Chen: "Measurement of redox states of mitochondrial cytochrome aa_3 in regions of liver loble by reflectance spectroscopy." Am.J.Physiol.264. G375-382 (1993)
• [Publications] T.Kinoshita: "Phosphorylation and dephosphorylation of guard-cell proteins from Vicia faba L.in response to light and dark." Plant Physiol.102. 917-923 (1993)
• [Publications] T.Fuse: "Characterization of a rice mutant having an increased susceptibility to light stress at high temperature." Physiol.Plantarum. 89. 799-804 (1993)
• [Publications] K.Iba: "A gene encoding a chloroplast ω-3 fatty acid desaturase complements alterations in fatty acid desaturataion and chloroplast copy number of the fad7 mutant of Arabidopsis thaliana." J.Biol.Chem.268. 24099-24105 (1993)
• [Publications] K.Kusumi: "Chlorophyll deficiency caused by a specific blocking of the C_5-pathway in virescent mutant seedlings of rice." Plant Cell Physiol.(in press). (1994)
• [Publications] K.Kusumi: "Regulation of PEP-carboxylase by biological clock in a CAM plant." Plant Cell Physiol.(in press). (1994)
• [Publications] N.Henomatsu: "Inhibition of intracellular transport of newly synthesized prolactin 6 by bafilomycin A1 in a pituitary tumor cell line,GH_3 cells." Eur.J.Cell Biol.62. 127-139 (1993)
• [Publications] T.Manabe: "Inhibitors of vacuolar-type H^+-ATPase suppresses proliferation of cultured cells." J.Cell Physiol.157. 445-452 (1993)
• [Publications] J.Deguchi: "Acidification of phagosomes and degradation of rod outer segments in rat retinal pigment epithelium." J.Ophotamol.Vis.Sci.35(in press). (1994)
• [Publications] Y.Moriyama: "ATP-dependent uptake of anti-neoplastic agents by acidic organelles." J.Biochem.(in press). (1994)
• [Publications] T.Okajima: "Site-directed random mutagenesis of AMP-binding residues in adenylate kinase." J.Biochem.114. 627-633 (1993)
• [Publications] T.Okajima: "Site-directed mutagenesis of AMP-binding residues in adenylate kinase:Alteration of substrate specificity." FEBS Lett.334. 86-88 (1993)
• [Publications] S.Kaya: "Pyridoxal-5'-phosphate probes at Lys-480 can sense the binding of ATP and the formation of phosphoenzymes in Na^+,K^+-ATPase." J.Biol.Chem.(in press). (1994)
• [Publications] Y.Mukohata: "Comparative studies on ion pumps of the bacteriorhodopsin family." Biophys.Chem.48(in press). (1994)
• [Publications] K.Ihara: "Met-145 is a key residue in the light-dark adaptation of bacterial rhodopsins." Biophys.J.(in press). (1994)
• [Publications] Y.Sugiyama: "Archaeopsin expressed in E.coli and its convertion to purple pigment in vitro." J.Biochem.(in press). (1994)
• [Publications] M.Tateno: "The proton pump rhodopsins from Haloarcula differ from bacteriorhodopsin and archaerhodopsins." Arch.Biochem.Biophys.(in press). (1994)
• [Publications] K.Saiki: "Identification of the functional domain in heme O synthase:Site-directed mutagenesis studies on the cyoE gene of the cytochrome bo operon in Escherichia coli." J.Biol.Chem.268. 26927-26934 (1993)
• [Publications] K.Saiki: "In vitro heme O synthesis by the cyoE gene product from Escherichia coli." J.Biol.Chem.268. 26041-26045 (1993)
• [Publications] M.Tsubaki: "Structure of the heme-copper binuclear center of the cytochrome bo complex of Escherichia coli:EPR and Fourier transform infrared spectroscopic studies." Biochemistry. 32. 6065-6072 (1993)
• [Publications] M.Tsubaki: "Cytochrome daxial ligand of the bd-type terminal quinol oxidase from Escherichia coli." FEBS Lett.335. 13-17 (1993)
• [Publications] T.Uno: "Resonance Raman and Fourier-transform infrared studies on the subunit I histidine mutants of the cytochrome bo complex in Escherichia coli:Molecular structure of redox metal centers." J.Biol.Chem.269(in press). (1994)
• [Publications] Y.N.Zhang: "Effects of mutagenic substitutions of prolines on the rate of deprotonation and reprotonation of the Schiff base during the photocycle of bacteriorhodopsin." Photochem.Photobiol.57. 1027-1031 (1993)
• [Publications] 佐伯和彦: "フェレドキシンの進化、はじめに" 新生化学実験講座 分子進化実験法(東京化学同人). 16. 153-154 (1993)
• [Publications] 香川靖雄: "酸化的リン酸化" 生体の科学 特集「現代医学、生物学の仮説、学説」(医学書院). 44. 410-411 (1993)
• [Publications] 香川靖雄: "細胞の一般生理" 標準生理学第3版(医学書院). 7-41 (1993)
• [Publications] 吉森保: "小胞輸送と選別の分子機構" 蛋白質核酸酵素 特集「分泌系蛋白質輸送の分子機構」. 38. 927-938 (1993)
• [Publications] 森山芳則: "液胞型H^+-ATPase阻害剤としてのバフィロマイシンおよび類縁化合物" 蛋白質核酸酵素 特集「細胞機能研究のための低分子プローブ」. 38. 2000-2011 (1993)
• [Publications] T.Fukui: "Molecular aspects of enzyme catalysis" Kodansha Ltd./VCH Verlags AG, 241 (1994)
• [Publications] 香川靖雄: "生体膜と疾患の分子生物学" 南山堂, 560 (1993)
• [Publications] 香川靖雄: "図説医化学第2版" 南山堂, 486 (1993)
• [Publications] 香川靖雄: "エネルギー、生命、環境そして栄養" 女子栄養大学出版部, 280 (1993)