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1991 Fiscal Year Final Research Report Summary

Molecular Science for Elucidation of Proton Active Transport and Electron Transfers through Proteins.

Research Project

Project/Area Number 02453157
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionOkazaki National Research Institutes, Institute for Molecular Science

Principal Investigator

KITAGAWA Teizo  Okazaki National Research Institutes Professor, 分子科学研究所, 教授 (40029955)

Co-Investigator(Kenkyū-buntansha) KAMOGAWA Keiji  Okazaki National Research Institutes Research Associate, 分子科学会究所, 助手 (40150057)
OGURA Takashi  Okazaki National Research Institutes Research Associate, 分子科学会究所, 助手 (70183770)
Project Period (FY) 1990 – 1991
KeywordsHemoglobin / Cytochrome c Oxidase / Time-resolved resonance Raman / UV resonance Raman / Proton Transport / Electron Transfer
Research Abstract

Active transport of protons by proteins is presumably based on pKa changes of some proton-carrying residues upon conformation change. In order to examine protonation-dependent protein dynamics, time-resolved ultraviolet resonance Raman spectra were pursued for the quaternary structure change of hemoglobin (Hb) from R to T upon photodissociation of its carbonmonoxide adduct. Raman spectra were probed by laser pulses at 218nm with 10ns width, while CO was photodissociated by another laser pulses at 416nm with lons width. The delay time of the probe pulse from the pump pulse was varied from -200mus to 500mus. The relative intensity of a tryptophane band at 1011cm^<-1> to a phenylalanine band at 1003cm^<-1> changed with the quaternary structure change. The change took place at 5 - 20mus after photolysis at pH7.4 but it had been completed at lOmus after photolysis at pH5.8 where Bohr residues are protonated. We deduced the tryptophane residue responsible for the Raman spectral change is beta37-Trp and discussed a mechanism by which a structural change of heme in alpha subunit is communicated to the beta subunit.
We also investigated reaction intermediates of cytochrome oxidase by using time-resolved resonance Raman spectroscopy and proposed likely structures of partially reduced oxygen. The results are published separately.

  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] S.Hashimoto,Y.Mizutani,Y.Tatsuno,T.Kitagawa: "Resonance Raman characterization of ferric-and ferrylporphyrin π cation radicals and the Fe^<IV>=O strerching frequency" J.Am.Chem.Soc.113. 6542-6549 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Egawa,T.Ogura,R.Makino,Y.Ishimura,T.Kitagawa: "Observation of the O-O stretching Raman band for cytochrome P-450_<cam> under catalytic conditions" J.Biol.Chem.266. 10246-10248 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Nagai,Y.Yoneyama,T.Kitagawa: "Unusual CO binding geometry in abnormal subunits of hemoglobin M Boston and Hb M Saskaton" Biochemistry. 30. 6495-6503 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Mizutani,S.Tokutomi,K.Aoyagi,K.Horitsu,T.Kitagawa: "Resonance Raman study on intact pea phytochrome and its model compounds;Evidence for proton migration during the phototransformation" Biochemistry. 30. 10693-10700 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Ogura,S.Takahashi,K.Shinzawa-Itoh,S,Yoshikawa,T.Kitagawa: "Time-resolved resonance Raman investigation of cytochrome oxidase catalysis:Observation of a new oxygen-isotope sensitive Raman band" Bull.Chem.Soc.Jpn.64. 2901-2907 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Takahashi,T.Ogura,K.Itoh-Shinzawa,S.Yoshikawa,T.Kitagawa: "Observation of the Cu_A-ligand stretching resonance Raman band for cytochrome c oxidase" J.Am.Chem.Soc.113. 9400-9401 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 北川 禎三: "ラマン分光,第4版 実験化学講座" 372-388 (1991)

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      「研究成果報告書概要(和文)」より
  • [Publications] 北川 禎三,紙中 庄司: "紫外共鳴ラマン分光による蛋白質の構造研究,分光研究" 193-206 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S. Sato, M. Asano-Someda, T. Kitagawa: "Time-resolved resonance Raman spectra of free-base octaethylporphyin in the S_1 and T_1 states" Chem. Phys. Lett.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Kaminaka, T. Kitagawa: "Time-resolved UV resonance Raman study on Bohr effects of hemoglobin" J. Am. Chem. Soc.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] "A new model for dioxygen binding in hemocyanin. Synthesis, characteri-zation, and molecular structure of the mu-eta^2 : eta^2 peroxo dinuclear copper (II) complexes, [Cu (HB (3, 5-R_2pz)_3) ]_2 (O_2) (R=iPr and Ph)" J. Am. Chem. Soc.

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Takahashi, T. Ogura, K. Itoh-Shinzawa, S. Yoshikawa, T. Kitagawa: "Observation of the Cu_A-ligand stretching resonance Raman band for cytochrome c oxidase" J. Am. Chem. Soc.113. 9400-9401 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N. Kitajima, T. Koda, S. Hashimoto, T. Kitagawa, Y. Moro-oka: "Synthesis and characterization of the dinuclear copper (II) complexes [Cu (HB (3, 5-Me_2pz)_3) ]_2 X (X=O^<2->, (OH)_2^<2->, CO_3^<2->, O_2^<2->)" J. Am. Chem. Soc.113. 5664-5671 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Ogura, S. Takahashi, K. Shinzawa-Itoh, S. Yoshikawa, T. Kitagawa: "Time-resolved resonance Raman investigation of cytochrome oxidase catalysis : Observation of a new oxygen-isotope sensitive Raman band" Bull. Chem. Soc. Jpn.64. 2901-2907 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Imai, K. Fushitani, G. Miyazaki, K. Ishimori, T. Kitagawa, Y. Wada, H. Morimoto, I. Morishima, D. T. -b Shih, J. Tame: "Site-directed mutagenesis in haemoglobin. Functional role of tyrosine-42 (C7) alpha at the alpha1-beta2" J. Mol. Biol.218. 769-778 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Mizutani, S. Tokutomi, K. Aoyagi, K. Horitsu, T. Kitagawa: "Resonance Raman study on intact pea phytochrome and its model compounds ; Evidence for proton migration during the phototransformation" Biochemistry. 30. 10693-10700 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M. Nagai, Y. Yoneyama, T. Kitagawa: "Unusual CO binding geometry in abnormal subunits of hemoglobin M Boston and Hb M Saskatoon" Biochemistry. 30. 6495-6503 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Egawa, T. Ogura, R. Makino, Y. Ishimura, T. Kitagawa: "Observation of the 0-0 stretching Raman band for cytochrome P-450_<cam> under catalytic conditions" J. Biol. Chem.266. 10246-10248 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Kamogawa, T. Kitagawa: "Evidence for direct intermolecular interactions as an origin of the hydration shifts of the C-H stretching vibrations : 1.4-dioxane / water system" Chem. Phys. Lett.179. 271-276 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] V. Fidler, T. Ogura, S. Sato, K. Aoyagi, T. Kitagawa: "Resonance Raman study on photoreduction of iron-porphyrins : a novel insight into the ligandaided process" Bull. Chem. Soc. Jpn.64. 2315-2322 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Hashimoto, Y. Mizutani, Y. Tatsuno, T. Kitagawa: "Resonance Raman characterization of ferric- and ferrylporphyrin pi cation radicals and the Fe^<IV>=0 stretching frequency" J. Am. Chem. Soc.113. 6542-6549 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Mizutani, K. Kamogawa, T. Kitagawa, A. Shimizu, Y. Taniguchi, K. Nakanishi: "Solution structures and phase separation in fluoroalcohol/water mixtures ; A study with Raman difference and ^<13>C NMR spectroscopy" J. Phys. Chem.95. 1790-1794 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M. Nakagawa, A. Maeda, T. Ogura, T. Kitagawa: "Identification of the long-lived L' species of bacteriorhodopsin to the N intermediate by Raman/absorption quasi-simultaneous measurements" J. Mol. Struct.242. 221-234 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Ogura, S. Takahashi, K. Shinzawa, S. Yoshikawa, T. Kitagawa: "Observation of the Fe^<II>-O_2 stretching Raman band for compound A of cytochrome oxidase at room temperature" J. Am. Chem. Soc.112. 5630-5631 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Mizutani, S. Hashimoto, Y. Tatsuno, T. Kitagawa: "Resonance Raman pursuit of the change from Fe^<II>-O_2 to Fe^<III>-OH via Fe^<IV>=O in autoxidation of ferrous iron-prophyrin" J. Am. Chem. Soc.112. 6809-6814 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Tokutomi, Y. Mizutani, E. Anni, T. Kitagawa: "Resonance Raman scattering from large pea phytochrome at ambient temperature : Difference in chromophore protonation between red- and far-red light absorbing forms" FEBS Lett.269. 341-344 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N. Kitajima, H. Fukui, Y. Moro-oka, Y. Mizutani, T. Kitagawa: "A synthetic model for dioxygen binding site of non-heme proteins. X-ray structure of Fe (OBz) (MeCN) [HB (3, 5-iPr_2pz)_3] and resonance Raman evidence for reversible formation of peroxo adduct" J. Am. Chem. Soc.112. 6402-6403 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Egawa, Y. Imai, T. Ogura, T. Kitagawa: "Resonance Raman study on mutant cytochrome P-450 obtained by site directed mutagenesis" Biochim. Biophys. Acta. 1040. 211-216 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Ogura, V. Fidler, Y. Ozaki, T. Kitagawa: "Alcohol catalyzed photoreduction of iron porphyrin complexes revealed by resonance Raman and absorption spectroscopies" Chem. Phys. Lett.169. 457-459 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Kamogawa, T. Kitagawa: "A new device for Raman difference spectroscopy and its application to observe frequency shifts due to isotope mixing" J. Phys. Chem.94. 3916-3921 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Kaminaka, T. Ogura, T. Kitagawa: "Time-resolved ultraviolet resonance Raman study of the photolysis of carbonmonoxy hemoglobin : Relaxation of the globin structure" J. Am. Chem. Soc.112. 23-27 (1990)

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      「研究成果報告書概要(欧文)」より

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Published: 1993-03-16  

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