2001 Fiscal Year Annual Research Report

生命1次情報の高速収集とそれに基づく統合バイオサイエンスの展開

Research Project

Project/Area Number	13GS0016
Research Category	Grant-in-Aid for Creative Basic Research
Research Institution	Okazaki National Research Institutes
Principal Investigator	永山國昭岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (70011731)
Co-Investigator(Kenkyū-buntansha)	岡村康司岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (80201987) 森泰生岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (80212265) 渡辺芳人岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (10201245) 高橋卓也岡崎国立共同研究機構, 計算科学研究センター, 助手 (70262102) 宇理須恒雄岡崎国立共同研究機構, 分子科学研究所, 教授 (50249950)
Keywords	電子顕微鏡 / 人工酵素 / カルシウム / チャネル / 蛋白質 / DNA / 信号制御 / 自己組織化
Research Abstract	生命一情報高速収集:1分子DNA配列直読法における要素技術(元素弁別電顕法、1本鎖DNAの延伸固定法、核酸塩基重元素ラベル法、特異的塩基対反応法、高速画像読取法)・膜蛋白質構造解析法における2次元結晶化法、蛋白質工学的電顕染色法における金属蛋白質の融合蛋白質発現手法等、の開発に着手した。電子顕微鏡用単結晶シリコン位相版のSOI基板に関して、SiとSiO2についての精密エッチング制御技術を確立し、極薄Si単結晶膜形成の準備を整えた。細胞系のin vivo生命情報:原索動物の胚発生における、局所Ca^<2+>濃度を制御する機能分子が集積する過程を解析し、Ca^<2+>イオンを正に制御する膜機能分子の発達が、負に制御する分子群の出現に先行することを見出した。また、細胞膜Ca^<2+>チャネル、細胞内膜Ca^<2+>放出チャネル、K^+チャネルの3者の空間的配置とCa^<2+>感受性による連携は、個体の運動の周波数である約15Hzの膜電位振動を生じることを明らかにした。細胞膜受容体活性化チャネルTRP1はストア依存性Ca^<2+>流入だけでなく、小胞体からのIP_3依存的Ca^<2+>放出をも制御することを示した。更には、細胞redox状態を感受し活性化して細胞死を仲介する、新規Ca^<2+>チャネルLTRPC2を同定した。生体シュミレーション:金属貯蔵蛋白質や膜チャネル蛋白質の機能メカニズムなど生理機能の調節機構を定量的に解明するため、鉄貯蔵蛋白質フェリチンフェリチン分子に関して、イオンの位置やアミノ酸残基置換の影響の実験データを、計算によって再現することができた。ナノバイオテクノロジーにおける応用:人工酵素による物質変換のシステム構築を目指し、シトクロームP450の機能を発現するヘム酵素の設計を行い、酸化活性種を与えるミオグロビンミュータントの反応サイトへ基質(トリプトファン)の導入を行い、ほぼ化学量論的に芳香環の水酸化反応を達成した。シリコンウエハー上の蛋白質集積パタン形成と新規バイオエレクトロニクス機能の研究においては、放射光ナノリソグラフィー専用のビームラインを建設し表面の平坦度Ra=0.33nmの超選択エッチングを確認した、また、Si(100)表面についてほぼ完全な水素終端表面を形成する、更に、その各種アルキル化合物との反応により、表面の親水性・疎水性を制御した自己組織単分子膜を形成する条件を見出した。

Research Products

(34 results)

All Other

All Publications (34 results)

[Publications] N.Makihara: "pH-Selective Hydrogenation of Water-soluble Carbon Compounds and Alkenes with [Cp*IrIII(H2O)3]2+(Cp*=h5C5H) as A catlyst Precusor in Very Acidic Media"Organometallics. 20. 497-500 (2001)
[Publications] H.Ohtsu: "Characterization of Imidazolate-Bridged Dinuclear and Mononuclear Hydroperoxo Complexes"Inort.Chem.. 40. 3200-3207 (2001)
[Publications] N.Haruta: "Protein Conformation Change of Myoglobin upon Ligand Binding Probed by Ultraviolet Resonance Raman Spectroscopy"Biochemistry. 40. 6956-6963 (2001)
[Publications] S.Herold: "Peroxynitrite Isomerization Catalyzed by His64 Myoglobin Mutants"J.Am.Chem.Soc.. 123. 4085-4086 (2001)
[Publications] T.Tomita: "Elucidation of the Differences between the 430 and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy"J.Biol.Chem.. 276. 36261-36267 (2001)
[Publications] I.Hara: "Oxidative Modification of Thyptophan-43 in the Heme Vicinity of the F43W/H64L Myoglobin Mutant"J.Biol.Chem.. 276. 36067-36070 (2001)
[Publications] Sandoz G.: "Distinct properties and differential β subunit regulation of two C-terminal isoforms of the P/Q-type Ca2+ channel α1A subunit"Eur.J.Neurosci.. 14. 987-997 (2001)
[Publications] R.Danev: "Transmission Electron Microsocpy with Zernike Phase Plate"Ultrarnicroscopy. 88. 243-252 (2001)
[Publications] Mori Y.: "Dissecting receptor-mediated Ca2+ influx pathways : TRP channels and their native counterparts"Jpn.J.Pharmacol. 87. 245-252 (2001)
[Publications] N.Takezoe: "Design and construction of UVSOR-BL4A2 beam line for nano-structure processing"Nucl.Instrum.Methods.. A467, Part2. 1279-1281 (2001)
[Publications] Y.Nonogaki: "STM observation of surface nanostructures on Si(111) formed after synchrotron radiation stimulated cleaning"Transactions MRS Japan. 26. 751-754 (2001)
[Publications] Y.Nonogaki: "Nanostructure formation on Si(111) surface assisted by synchrotron radiation illumination-Characterization by scanning tunneling microscopy"J.Electron spectroscopy. 119. 241-246 (2001)
[Publications] S.Ogo: "pH-Dependent Transfer Hydrogenation, Reductive Amination, and Dehalogenation of Water-Soluble Carbonyl Compounds and Alkyl halides Promoted by CP*lr Complexes"Organomatellics. 20. 4903-4910 (2001)
[Publications] S.Ogo: "pH-Dependent H2-Activation Cycle to Reducection of Nitrate Ion by Metal Complexes"J.Am.Chem.Soc.. 124. 597-601 (2002)
[Publications] Y.Watanabe: "Asymmetric Oxidation of Sulfides and olefins by Myoglobin Mutants"Rev.Heteroatom Chem.. 22. 135-150 (2001)
[Publications] J.L.Que: "Oxygenase Pathways : Oxo, Peroxo, and Superoxo"Science. 292. 651-653 (2001)
[Publications] S.Ozaki: "Investigations of the Roles of the Distal Heme Environment and the Proximal Heme Iron Ligand in Peroxide Activation by Heme Enzymes via Molecular Engineering of Myoglobin"Acc.Chem.Res.. 34. 818-825 (2001)
[Publications] Y.Watanabe: "Two-Electron Oxidized Derivatives of Fe (III) Porphyrins"J.Biol.Inorg.Chem.. 6. 846-856 (2001)
[Publications] Okada, T.: "The form of neurons in the ascidian central nervous system of the tadpole larva of Ciona"Biol.Bull.. 200. 252-256 (2001)
[Publications] Ohtsuka, Y.: "Changes in gelsolin expression during ascidian metamorphosis"Development Gene & Evolution. 211. 252-256 (2001)
[Publications] K.Danov: "Electric charging of thin films measured using the constrast transfer function"Ultramicroscopy. 87. 45-54 (2001)
[Publications] D.Tanooka: "Color-imaging ellipsometer : High-Seed characterization of in-plane distribution of film thickness at nano-scale"Jpn.J.Appl.Phys.. 40. 877-880 (2001)
[Publications] R.Danev: "Complex Observation in Electron Microscopy. II. Direct Visualization Phases and Amplitudes of Exit Wave Functions"J.Phys.Soc.Jpn.. 70. 696-702 (2001)
[Publications] K.Murata: "Asymmetric arrangement of auxiliary subunits of skeletal muscle voltage-gated L-type Ca2+ channel"Biochim.Biophys.Res.Commun.. 282. 284-291 (2001)
[Publications] Ino M.: "Functional disorders of the sympathetic nervous system in mice lacking the α 1B subunit (Cav 2.2) of N-tupe calclum channel"Proc.Natl.Acad.Sci.USA. 98. 5323-5328 (2001)
[Publications] Kato Y.: "Evidence in support of a four transmembrane-pore-transmembrane topolpgy model for the Arabidopsis thaliana Na+/K+ translocation AtHKT1 protein, a member of the superfamily of K+ transporters"Proc.Natl.Acad.Sci.USA. 98. 6488-6493 (2001)
[Publications] Hatakeyama S.: "Differential nociceptive responses in mice lacking subunit the α 1B subunit of N-type Ca2+ channels"Neuroreport. 12. 2423-2427 (2001)
[Publications] Kinoshita M.: "Binding of G α 0 N-terminus is responsible for the voltage-resisitant inhibition of α 1B (P/Q-type, Cav 2.1) Ca2+ channels"J.Biol.Chem.. 276. 28731-28783 (2001)
[Publications] Kang MG.: "Biochemical and biophysical evidence for γ2 subunit association with neuronal voltaae-actlvated Ca2+ channels"J.Biol.Chem.. 276. 32917-32924 (2001)
[Publications] Kanki H.: "Activation of inisitol 1, 4, 5-trlsphosphate receptor is essential for the opening of mouse TRP5 channels"Mol.Pharmacol.. 60. 989-998 (2001)
[Publications] Ohtsuka, Y: "Induction of ascidian peripheral neuron vegetal blastomeres"Dev.Biol.. 239(1). 107-117 (2001)
[Publications] Murata, Y.: "Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate"Receptors and Channels. 7. 387-399 (2001)
[Publications] Ohtsuka, Y.: "Induction of ascidian peripharal neuron by vegetal blastomeres"Dev.Biol.. 239. 107-117 (2001)
[Publications] Meinertzhagen, I.A.: "The ascidian larval nervous system : the chordate brain from its small beginnings"Trends in Neurosci.. 24(7). 401-410 (2001)

2001 Fiscal Year Annual Research Report

生命1次情報の高速収集とそれに基づく統合バイオサイエンスの展開

Principal Investigator

永山 國昭 岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (70011731)

Research Products

[Publications] N.Makihara: "pH-Selective Hydrogenation of Water-soluble Carbon Compounds and Alkenes with [Cp*IrIII(H2O)3]2+(Cp*=h5C5H) as A catlyst Precusor in Very Acidic Media"Organometallics. 20. 497-500 (2001)

[Publications] H.Ohtsu: "Characterization of Imidazolate-Bridged Dinuclear and Mononuclear Hydroperoxo Complexes"Inort.Chem.. 40. 3200-3207 (2001)

[Publications] N.Haruta: "Protein Conformation Change of Myoglobin upon Ligand Binding Probed by Ultraviolet Resonance Raman Spectroscopy"Biochemistry. 40. 6956-6963 (2001)

[Publications] S.Herold: "Peroxynitrite Isomerization Catalyzed by His64 Myoglobin Mutants"J.Am.Chem.Soc.. 123. 4085-4086 (2001)

[Publications] T.Tomita: "Elucidation of the Differences between the 430 and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy"J.Biol.Chem.. 276. 36261-36267 (2001)

[Publications] I.Hara: "Oxidative Modification of Thyptophan-43 in the Heme Vicinity of the F43W/H64L Myoglobin Mutant"J.Biol.Chem.. 276. 36067-36070 (2001)

[Publications] Sandoz G.: "Distinct properties and differential β subunit regulation of two C-terminal isoforms of the P/Q-type Ca2+ channel α1A subunit"Eur.J.Neurosci.. 14. 987-997 (2001)

[Publications] R.Danev: "Transmission Electron Microsocpy with Zernike Phase Plate"Ultrarnicroscopy. 88. 243-252 (2001)

[Publications] Mori Y.: "Dissecting receptor-mediated Ca2+ influx pathways : TRP channels and their native counterparts"Jpn.J.Pharmacol. 87. 245-252 (2001)

[Publications] N.Takezoe: "Design and construction of UVSOR-BL4A2 beam line for nano-structure processing"Nucl.Instrum.Methods.. A467, Part2. 1279-1281 (2001)

[Publications] Y.Nonogaki: "STM observation of surface nanostructures on Si(111) formed after synchrotron radiation stimulated cleaning"Transactions MRS Japan. 26. 751-754 (2001)

[Publications] Y.Nonogaki: "Nanostructure formation on Si(111) surface assisted by synchrotron radiation illumination-Characterization by scanning tunneling microscopy"J.Electron spectroscopy. 119. 241-246 (2001)

[Publications] S.Ogo: "pH-Dependent Transfer Hydrogenation, Reductive Amination, and Dehalogenation of Water-Soluble Carbonyl Compounds and Alkyl halides Promoted by CP*lr Complexes"Organomatellics. 20. 4903-4910 (2001)

[Publications] S.Ogo: "pH-Dependent H2-Activation Cycle to Reducection of Nitrate Ion by Metal Complexes"J.Am.Chem.Soc.. 124. 597-601 (2002)

[Publications] Y.Watanabe: "Asymmetric Oxidation of Sulfides and olefins by Myoglobin Mutants"Rev.Heteroatom Chem.. 22. 135-150 (2001)

[Publications] J.L.Que: "Oxygenase Pathways : Oxo, Peroxo, and Superoxo"Science. 292. 651-653 (2001)

[Publications] S.Ozaki: "Investigations of the Roles of the Distal Heme Environment and the Proximal Heme Iron Ligand in Peroxide Activation by Heme Enzymes via Molecular Engineering of Myoglobin"Acc.Chem.Res.. 34. 818-825 (2001)

[Publications] Y.Watanabe: "Two-Electron Oxidized Derivatives of Fe (III) Porphyrins"J.Biol.Inorg.Chem.. 6. 846-856 (2001)

[Publications] Okada, T.: "The form of neurons in the ascidian central nervous system of the tadpole larva of Ciona"Biol.Bull.. 200. 252-256 (2001)

[Publications] Ohtsuka, Y.: "Changes in gelsolin expression during ascidian metamorphosis"Development Gene & Evolution. 211. 252-256 (2001)

[Publications] K.Danov: "Electric charging of thin films measured using the constrast transfer function"Ultramicroscopy. 87. 45-54 (2001)

[Publications] D.Tanooka: "Color-imaging ellipsometer : High-Seed characterization of in-plane distribution of film thickness at nano-scale"Jpn.J.Appl.Phys.. 40. 877-880 (2001)

[Publications] R.Danev: "Complex Observation in Electron Microscopy. II. Direct Visualization Phases and Amplitudes of Exit Wave Functions"J.Phys.Soc.Jpn.. 70. 696-702 (2001)

[Publications] K.Murata: "Asymmetric arrangement of auxiliary subunits of skeletal muscle voltage-gated L-type Ca2+ channel"Biochim.Biophys.Res.Commun.. 282. 284-291 (2001)

[Publications] Ino M.: "Functional disorders of the sympathetic nervous system in mice lacking the α 1B subunit (Cav 2.2) of N-tupe calclum channel"Proc.Natl.Acad.Sci.USA. 98. 5323-5328 (2001)

[Publications] Kato Y.: "Evidence in support of a four transmembrane-pore-transmembrane topolpgy model for the Arabidopsis thaliana Na+/K+ translocation AtHKT1 protein, a member of the superfamily of K+ transporters"Proc.Natl.Acad.Sci.USA. 98. 6488-6493 (2001)

[Publications] Hatakeyama S.: "Differential nociceptive responses in mice lacking subunit the α 1B subunit of N-type Ca2+ channels"Neuroreport. 12. 2423-2427 (2001)

[Publications] Kinoshita M.: "Binding of G α 0 N-terminus is responsible for the voltage-resisitant inhibition of α 1B (P/Q-type, Cav 2.1) Ca2+ channels"J.Biol.Chem.. 276. 28731-28783 (2001)

[Publications] Kang MG.: "Biochemical and biophysical evidence for γ2 subunit association with neuronal voltaae-actlvated Ca2+ channels"J.Biol.Chem.. 276. 32917-32924 (2001)

[Publications] Kanki H.: "Activation of inisitol 1, 4, 5-trlsphosphate receptor is essential for the opening of mouse TRP5 channels"Mol.Pharmacol.. 60. 989-998 (2001)

[Publications] Ohtsuka, Y: "Induction of ascidian peripheral neuron vegetal blastomeres"Dev.Biol.. 239(1). 107-117 (2001)

[Publications] Murata, Y.: "Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate"Receptors and Channels. 7. 387-399 (2001)

[Publications] Ohtsuka, Y.: "Induction of ascidian peripharal neuron by vegetal blastomeres"Dev.Biol.. 239. 107-117 (2001)

[Publications] Meinertzhagen, I.A.: "The ascidian larval nervous system : the chordate brain from its small beginnings"Trends in Neurosci.. 24(7). 401-410 (2001)

永山國昭岡崎国立共同研究機構, 統合バイオサイエンスセンター, 教授 (70011731)

[Publications] N.Makihara: "pH-Selective Hydrogenation of Water-soluble Carbon Compounds and Alkenes with [CpIrIII(H2O)3]2+(Cp=h5C5H) as A catlyst Precusor in Very Acidic Media"Organometallics. 20. 497-500 (2001)