1995 Fiscal Year Annual Research Report

酸素結合蛋白質の分子操作と精密構造機能研究の新展開

Research Project

Project/Area Number	07308071
Research Category	Grant-in-Aid for Co-operative Research (A)
Research Institution	Osaka University
Principal Investigator	今井清博大阪大学, 医学部, 助教授 (50028528)
Co-Investigator(Kenkyū-buntansha)	長井雅子金沢大学, 医学部, 教授 (60019578) 北川禎三岡崎国立共同研究機構, 分子科学研究所, 教授 (40029955) 飯塚哲太郎理化学研究所, 主任研究員 (30029475) 森島績京都大学, 工学部, 教授 (50026093) 森本英樹大阪大学, 基礎工学部, 助教授 (20029474)
Keywords	酸素結合蛋白質 / ヘモグロビン / ミオグロビン / 人工変異 / 分子分光学 / 分子進化 / 分子動力学
Research Abstract	種々の酸素結合蛋白質の機能発現分子機構の研究の新展開をめざし、以下の結果を得た。 1.ウシ・ヘモグロビンの真性低酸素親和性を模擬するための3種類の組換えヘモグロビンの酸素平衡(今井)、α鎖、β鎖、ミオグロビンのモジュール同士を組み換えた9種類のキメラグロビンのヘム結合能、二次構造形成能などの解析(森島)、Mg-Fe混成ヘモグロビンとその人工変異体(βEllVal→Ile)のX線結晶解析によるヘムとその近傍の微細な構造変化の検出(森本) 2.モノアザヘムまたはジアザヘムをもつ還元型再構成ミオグロビンにはピリジン、イミダゾール、シアン、アジドが安定に結合する(根矢)。 3.T状態マーカーとをしての近紫外域の負のCD帯やTyr残基に帰属する紫外域共鳴ラマン線を用いたHb M Bostonにおける構造転移の観測(長井)。 4.ナノ秒時間分解共鳴ラマン分光法によって求めた種々の遠位側アミノ酸残基置換型COMbの光解離後再結合過程でのFe-co伸縮振動数は、再結合が遅いほど高くなる(北川)。 5.ストップドフローX線散乱法によって蛋白質のフォールディングを追跡すると、ヘムの誘導によってミオグロビンがフォールドする条件を選べることが分かった(木原)。 6.シトクロムb558のスーパーオキサイド産生はヘム端経由での1電子移行によって起こる(飯塚)。 7.環形動物多毛類の巨大ヘモグロビン分子の階層的構造を安定化するリンカー鎖にレクチン類との相同性がみられ、そのレクチン活性が糖鎖を介する分子骨核形成に役立っている(後藤)。 8.インドールアミン酸素添加酵素から進化したミオグロビンはトコブシやアワビ以外のサザエやバテイラでも発見され、それらの一次構造のホモロジーを検討した(鈴木)。 9.低温のミオグロビン結晶のX線回析強度データに対して基準振動精密化法を適用し、蛋白質の温度因子のみならず各基準振動モードの温度依存性に関する情報を得た(郷)。

Research Products

(21 results)

All Other

All Publications (21 results)

[Publications] K.Imai: "Oxygen brinding properties of Eudistylia vancouverii chlorocruorin and its dodecamer subunit." Comp.Biochem.Physiol.(in press).
[Publications] T.Harano: "Hb Kurosaki[α7(A5)Lys→Glu]:A new α chain variant found in a Japanese women." Hemoglobin. 19. 197-201 (1995)
[Publications] S-Y.Park: "High resolution crystal structure of magnesium(Mg II)-iron(Fe II)hybrid hemoglobin with liganded beta subunits." J.Mol.Biol.255. 726-734 (1996)
[Publications] N.Shibayama: "Oxygen equilibrium properties of nikel(II)-iron(II)hybrid hemoglobins cross-linked between 82 β land 82 β 2 lysyl residues by bis(3,5-dibromosalicyl)fumarate:Determination of the first two-step microscopic Adair constants for human hemoglobin." Biochemistry. 34. 4773-4780 (1995)
[Publications] K.Wakasugi: "NMR Studies on Recombinant Cytochrome P450cam Mutants." Biochimie. (in press). (1996)
[Publications] Y.Isogai: "The Mechanism of electron donation to molecular oxygen by phagocytic cytochrome b558" J.Biol.Chem. 270. 7853-7857 (1995)
[Publications] Y.Shiro: "Structure and function of nitric oxide reducing heme-enzyme cytochrome P450nor." J.Inorg.Biochem.59. 440 (1995)
[Publications] P.Jewsbury: "Distal residue-CO interaction in carbonmonoxy myoglobins : A molecular dynamics study of three distal mutants." Biophy.J. 68. 1283-1294 (1995)
[Publications] P.Jewsbury: "The proximal residue largely determines the CO distortion in carbon monoxy globin protein an ab initio study of a heme prosthetic unit." J.Phys.Chem.99. 12677-12685 (1995)
[Publications] M.Nagai: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan β 37 mutant." J.Biol.Chem.270. 1636-1642 (1995)
[Publications] M.Nagai: "UV resonance Raman studies of quaternary structure changes of human hemoglobin using a tyrosine α 42 mutant." J.Inorg.Biochem.59. 347 (1995)
[Publications] M.Tomimoto: "Analytic theory of pseudorotation in five-membered rings.Cyclopentane,tetrahydrofuran,ribose and deoxyribose." J.Phys.Chem.99. 563-577 (1995)
[Publications] S.Hayward: "Harmonicity and anharmonicity in protein dynamics : A normal mode analysis and principal component analysis." Proteins. 232. 177-186 (1995)
[Publications] Y.Igarashi: "Solution x-ray scattering study on the chaperonin GroEL for Escherichia coli." Biopys.Chem.53. 259-266 (1995)
[Publications] I.Narumi: "Bacillus stearothermophileus plasmid pSTK1 replicon is functional in Escherichia coli." Biotech.Lett.17. 475-480 (1995)
[Publications] S.Neya: "Novel ligand binding properties of the myoglobin reconstituted with monoazahemin." J.Biol.Chem.270. 13118-13123 (1995)
[Publications] 根矢三郎: "ミオグロビン内部のヘムのダイナミクス" 蛋白質核酸酵素. 40. 1115-1121 (1995)
[Publications] S.Ebina: "Carbohydrate gluing,an architectural mechanism in the supramolecular structure of an annelid giant hemoglobin." Proc.Natl.Acad.Sci.USA. 92. 7367-7371 (1995)
[Publications] T.Gotoh: "Disasscmbly and reassembly of giant hemoglobin from the polychaete Perinereis aibuhitensis" Physiol.Zool.68. 81 (1995)
[Publications] T.Suzuki: "Primary structure of a constituent polypeptide chain of the chlorocruorin from Sabellastarte indica." Biochim.Biophys.Acta. 1252. 189-193 (1995)
[Publications] T Suzuki: "Two-domain hemoglobin of the blood clam Barbatia lima resulted from the recent gene duplication of the single-domain δ chain." Biochem.J.313. 561-566 (1996)

1995 Fiscal Year Annual Research Report

酸素結合蛋白質の分子操作と精密構造機能研究の新展開

Principal Investigator

今井 清博 大阪大学, 医学部, 助教授 (50028528)

Research Products

[Publications] K.Imai: "Oxygen brinding properties of Eudistylia vancouverii chlorocruorin and its dodecamer subunit." Comp.Biochem.Physiol.(in press).

[Publications] T.Harano: "Hb Kurosaki[α7(A5)Lys→Glu]:A new α chain variant found in a Japanese women." Hemoglobin. 19. 197-201 (1995)

[Publications] S-Y.Park: "High resolution crystal structure of magnesium(Mg II)-iron(Fe II)hybrid hemoglobin with liganded beta subunits." J.Mol.Biol.255. 726-734 (1996)

[Publications] K.Wakasugi: "NMR Studies on Recombinant Cytochrome P450cam Mutants." Biochimie. (in press). (1996)

[Publications] Y.Isogai: "The Mechanism of electron donation to molecular oxygen by phagocytic cytochrome b558" J.Biol.Chem. 270. 7853-7857 (1995)

[Publications] Y.Shiro: "Structure and function of nitric oxide reducing heme-enzyme cytochrome P450nor." J.Inorg.Biochem.59. 440 (1995)

[Publications] P.Jewsbury: "Distal residue-CO interaction in carbonmonoxy myoglobins : A molecular dynamics study of three distal mutants." Biophy.J. 68. 1283-1294 (1995)

[Publications] P.Jewsbury: "The proximal residue largely determines the CO distortion in carbon monoxy globin protein an ab initio study of a heme prosthetic unit." J.Phys.Chem.99. 12677-12685 (1995)

[Publications] M.Nagai: "Ultraviolet resonance Raman studies of quaternary structure of hemoglobin using a tryptophan β 37 mutant." J.Biol.Chem.270. 1636-1642 (1995)

[Publications] M.Nagai: "UV resonance Raman studies of quaternary structure changes of human hemoglobin using a tyrosine α 42 mutant." J.Inorg.Biochem.59. 347 (1995)

[Publications] M.Tomimoto: "Analytic theory of pseudorotation in five-membered rings.Cyclopentane,tetrahydrofuran,ribose and deoxyribose." J.Phys.Chem.99. 563-577 (1995)

[Publications] S.Hayward: "Harmonicity and anharmonicity in protein dynamics : A normal mode analysis and principal component analysis." Proteins. 232. 177-186 (1995)

[Publications] Y.Igarashi: "Solution x-ray scattering study on the chaperonin GroEL for Escherichia coli." Biopys.Chem.53. 259-266 (1995)

[Publications] I.Narumi: "Bacillus stearothermophileus plasmid pSTK1 replicon is functional in Escherichia coli." Biotech.Lett.17. 475-480 (1995)

[Publications] S.Neya: "Novel ligand binding properties of the myoglobin reconstituted with monoazahemin." J.Biol.Chem.270. 13118-13123 (1995)

[Publications] 根矢 三郎: "ミオグロビン内部のヘムのダイナミクス" 蛋白質核酸酵素. 40. 1115-1121 (1995)

[Publications] S.Ebina: "Carbohydrate gluing,an architectural mechanism in the supramolecular structure of an annelid giant hemoglobin." Proc.Natl.Acad.Sci.USA. 92. 7367-7371 (1995)

[Publications] T.Gotoh: "Disasscmbly and reassembly of giant hemoglobin from the polychaete Perinereis aibuhitensis" Physiol.Zool.68. 81 (1995)

[Publications] T.Suzuki: "Primary structure of a constituent polypeptide chain of the chlorocruorin from Sabellastarte indica." Biochim.Biophys.Acta. 1252. 189-193 (1995)

[Publications] T Suzuki: "Two-domain hemoglobin of the blood clam Barbatia lima resulted from the recent gene duplication of the single-domain δ chain." Biochem.J.313. 561-566 (1996)

今井清博大阪大学, 医学部, 助教授 (50028528)

[Publications] 根矢三郎: "ミオグロビン内部のヘムのダイナミクス" 蛋白質核酸酵素. 40. 1115-1121 (1995)