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Development of high performance photo-diode-array spectrophotometer using weak probe light

Research Project

Project/Area Number 09354010
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field 動物生理・代謝
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

SHICHIDA Yoshinori  Kyoto University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (60127090)

Co-Investigator(Kenkyū-buntansha) SATO Tatsumi  Shimadzu Corporation, Analytical Instruments Division, Product Manager, 分析機器事業部, 課長
佐藤 辰己  (株)島津製作所, 分析機器事業部・技術部, プロダクトマネージャ
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥14,100,000 (Direct Cost: ¥14,100,000)
Fiscal Year 1998: ¥4,200,000 (Direct Cost: ¥4,200,000)
Fiscal Year 1997: ¥9,900,000 (Direct Cost: ¥9,900,000)
KeywordsSpectroscopy / Photo-diode-array / Spectrophotometer / Reaetion kinetics / Rhodopsin / Transducin / G-protein / Intermediate / ロドプシン / 光 / タンパク質 / 光反応 / 生体試料 / 高速 / 低温
Research Abstract

Visual pigment is a prototypical G-protein coupled receptor that receives a light signal from the environment using a light-absorbing chromophore, 11-cis-retinal Recently, we have shown that the low-temperature time-resolved spectroscopy is one of the powerful tools for elucidating the conformational change of visual pigments by monitoring change in absorption spectra. However, low time-resolution (-min) of a scanning type spectrophotometer hampered us from complete detection of change in absorption spectra after the irradiation.
In this project, we have developed a unique photo-diode-array (PDA) spectrophotometer to measure the spectral change with higher time-resolution (-sec). For this purpose, intensity of probe light is reduced with a electric shutter system and a glass band-pass filter, to suppress the photoreaction of the sample during the recording of the spectra. A sample cell holder connected to a thermostatic circulator or a optical cryostat was installed into the sample comp … More artment of the spectrophotometer to keep the sample at a constant temperature. Specially designed optical setup including shutters and a movable mirrors are equipped to the compartment to irradiate the sample in the optical cell without removing from cell holder. Using this spectrophotometer, we were able to investigate thermal reactions of meta-intermediates of bovine rhodopsin with time-resolution of 1 sec.
Irradiation of rhodopsin sample at - 10 。C caused formation of meta I-intermediate. Transition from meta I-intermediate to meta II was observed with time constant of -10 s which could not be detected with the scanning type spectrophotometer at the temperature. Addition of retinal G-protein, transducin caused accumulation of meta II, suggesting that meta II is a intermediate which interact with transducin. The accumulation process was also observed with time constant of -10 s. Thus, this unique PDA spectrophotometer is a powerful tool for recording change in absorption spectra of biological sample with time-resolution of -s. Less

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (65 results)

All Other

All Publications (65 results)

  • [Publications] H.Imai: "Single amino acid residue as a functional determinant of rod and cone pigments." Proc.Natl.Acad.Sci.USA. 94. 2322-2326 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Nagata: "Water and peptide bockborn structure in the active center of bovine rhodopsin." Biochemistry. 36. 6164-6170 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Imai: "Photochemical and biochemical propertes of chicken blue-sensitive cone visual pigment." Biochemistry. 36. 12773-12779 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] D.Kojima: "A novel Go-mediated phototransduction cascade in scallop visual cells." J.Biol.Chem.272. 22979-22982 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Tachibanaki: "Presence of two rhodopsin intermediates responsible for transducin activation." Biochemistry. 36. 14173-14180 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 今井啓雄: "時間分解低温分光法による蛋白質構造変化の解析" 生物物理. 37. 225-226 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] R.Akiyama: "Analysis of excited state dynamics of 13-trans-locked-bacteriorhodopsin." J.Phys.Chem.101. 412-417 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Wada: "Retinoids and related compounds. Part 20. Synthesis of 11-Z-8, 18-ethanoretinal and conformational study of rhodopsin chromophore." Chem.Soc.Perkin.Trans.1. 1773-1777 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Nagata: "The hydrogen bonding network of water molecules and the peptide backbone in the region connecting Asp83, Gly120 and Glu113 in bovin rhodopsin." Biochemistry. 37. 17216-17222 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Terakita: "Selective activation of G-protein subtypes by vertebrate and invertebrate rhodopsins." FEBS Lett.439. 110-114 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Wada: "Stereoselective synthesis of 11Z-9-demethyl-9-benzyl-and 9-phenyl-retinals and their interaction with bovine opsin." Bioorg.Med.Chem.Lett.89. 423-426 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Shichida: "Visual pigment : G-protein-coupled receptor for light signal." Cell Mol Life Sci.54. 1299-1315 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phys.Chem.107. 1334-1339 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Kandori: "Cysteine S-H as a hydrogen-bonding probe in proteins." J. Am. Chem. Soc.120. 5828-5829 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activates transducin in the bleaching process of bovine rhodopsin." FEBS Lett.425. 126-130 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Kandori: "Protein structural changes in bacteriorhodopsin upon photoisomerization as revealed by polarized FTIR spectroscopy" J. Phys. Chem.102. 7899-7905 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Chosrowjan: "Rhodopsin emission in real time : A new aspect of the primary event in vision." J. Am. Chem. Soc.120. 9706-9707 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 寺北明久: "視物質と視細胞光情報伝達系の多様性-機能解析と分子系統解析の接点" 蛋白質・核酸・酵素. 44. 217-225 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 七田芳則: "生物物理から見た生命像4 視る感覚「光のセンサー ; 視物質」(分担執筆)" 吉岡書店, 127 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 七田芳則: "視覚、図で学び絵で見るレーザー科学入門(分担執筆)" 丸善, (CD-ROM) (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hiroc Imai: "Analysis of Amino Acid Residuesin Rhodopsin and Cone Visual Pigments that Determine their Molecular Properties. (Methods in Enrynology 分担執筆)" Academic Press 印刷中, (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yoshinori Shichida: "Heterogeniety of Rhodopsin Intermediate State Interacting with Transducin. (Methods in Enrynology 分担執筆)" Academic Press 印刷中, (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Shichida: "Visual pigment : Photochemistry and molecular evolution.in The Retinal Basis of Vision. (ed.J.Toyoda)" Elsevier Science, The Netherlands. in press,

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] R.Akiyama: "Analysis of excited state dynamics of 13-trans-locked-bacteriorhodopsin." J.Phys.Chem.101. 412-417 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Imai: "Single amino acid residue as a functional determinant of rod and cone pigments." Proc.Natl.Acad.Sci.USA. 94. 2322-2326 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Nagata: "Water and peptide backbone structure in the active center of bovine rhodopsin." Biochemistry. 36. 6164-6170 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Wada: "Retinoids and related compounds. Part 20. Synthesis of 11-Z-8,18-ethanoretinal and conformational study of rhodopsin chromophore." Chem.Soc.Perkin Trans.1. 1773-1777 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Imai: "Photochemical and biochemical properties of chicken blue-sensitive cone visual pigment." Biochemistry. 36. 12773-12779 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] D.Kojima: "A novel Go-mediated phototransduction cascade in scallop visual cells." J.Biol.Chem.272. 22979-22982 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Tachibanaki: "Presence of two rhodopsin intermediates responsible for transducin activation." Biochemistry. 36. 14173-14180 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phys.Chem.107. 1334-1339

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Kandori: "Cysteine S-H as a hydrogen-bonding probe in proteins." J.Am.Chem.Soc.120. 5828-5829

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activates transducin in the bleaching process of bovine rhodopsin." FEBS Lett.425. 126-130

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Kandori: "Protein structural changes in bacteriorhodopsin upon photoisomerization as revealed by polarized FTIR spectroscopy." J.Phys.Chem.102. 7899-7905

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] H.Chosrowjan: "Rhodopsin emission in real time : A new aspect of the primary event in vision." J.Am.Chem.Soc.120. 9706-9707

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Nagata: "The hydrogen bonding network of water molecules and the peptide backbone in the region connecting Asp83, Gly120 and Glu113 in bovine rhodopsin." Biochemistry. 37. 17216-17222

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Terakita: "Selective activation of G-protein subtypes by vertebrate and invertebrate rhodopsins." FEBS Lett.439. 110-114

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] A.Wada: "Stereoselective synthesis of 11Z-9-demethyl-9-benzyl-and 9-phenyl-retinals and their interaction with bovine opsin." Bioorg.Med.Chem.Lett.8. 423-426

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Y.Shichida: "Visual Pigment : G-protein-coupled receptor for light signal." Cell Mol Life Sci.54. 1299-1315

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phys.Chem.107. 1334-1339 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Kandori: "Cysteine S-H as a hydrogen-bonding probe in proteins." J.Am.Chem.Soc.120. 5828-5829 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activates transducin in the bleaching process of bovine rhodopsin." FEBS Lett.425. 126-130 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Kandori: "Protein structural changes in bacteriorhodopsin upon photoisomerization as revealed by polarized FTIR spectroscopy." J.Phys.Chem.102. 7899-7905 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Chosrowjan: "Rhodopsin emission in real time: A new aspect of the primary event in vision." J.Am.Chem.Soc.120. 9706-9707 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] T.Nagata: "The hydrogen bonding network of water molecules and the peptide backbone in the region connecting Asp83,Gly120 and Glu113 in bovine rhodopsin." Biochemistry. 37. 17216-17222 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Terakita: "Selective activation of G-protein subtypes by vertebrate and invertebrate rhodopsins." FEBS Lett.439. 110-114 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] A.Wada: "Stereoselective synthesis of 11Z-9-demethyl-9-benzyl-and 9-phenyl-retinals and their interaction with bovine opsin." Bioorg.Med.Chem.Lett.8. 423-426 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Shichida: "Visual Pigment: G-protein-coupled receptor for light signal." Cell Mol Life Sci.54. 1299-1315 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 寺北明久: "視物質と視細胞光情報伝達系の多様性-機能解析と分子系統解析の接点" 蛋白質・核酸・酵素. 44. 217-225 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hiroo Imai: "Analysis of Amino Acid Residuesin Rhodopsin and Cone Visual Pigments that Determine their Molecular Properties. (Methods in Enzynology 分担執筆)" Academic Press(印刷中), (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yoshinori Shichida: "Heterogeniety of Rhodopsin Intermediate State Interacting with Transducin.(Methods in Enzynology 分担執筆)" Academic Press(印刷中), (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Y.Shichida: "Visual pigment: Photochemistry and molecular evolution.in The Retinal Basis of Vision.(ed.J.Toyoda)" Elsevier Science,The Netherlands.(in press),

    • Related Report
      1998 Annual Research Report
  • [Publications] H.Imai: "Single amino acid residue as a functional determinant of rod and cone pigments." Proc.Natl.Acad.Sci.USA. 94. 2322-2326 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Nagata: "Water and peptide bockborn structure in the active center of bovine rhodopsin." Biochemistry. 36. 6164-6170 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Imai: "Photochemical and biochemical properties of chicken blue-sensitive cone visual pigment." Biochemistry. 36. 12773-12779 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] D.Kojima: "A novel Go-mediated phototransduction cascade in scallop visual cells." J.Bio.Chem.272. 22979-22982 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Tachibanaki: "Presence of two rhodopsin intermediates responsible for transducin activation." Biochemistry. 36. 14173-14180 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] S.Tachibanaki: "Identification of a new intermediate state that binds but not activate transducin in the bleaching process of bovine rhodopsin." FEBS lett.(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] R.Akiyama: "Analysis of excited state dynamics of 13-trans-locked-bacteriorhodopsin." J.Phys.Chem.101. 412-417 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] A.Wada: "Retinoids and related compounds.Part 20.Synthesis of 11-Z-8,18-ethanoretinal and conformational study of rhodopsin chromophore." Chem.Soc.Perkin Trans.1. 1773-1777 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Kakitani: "Deuterium substitution effect on the excited state dynamics of rhodopsin." J.Phy.Chem.(印刷中). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Shichida: "Visual Pigments." J.Cell,Mol.Life Sci.(印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] 今井啓雄: "時間分解低温分光法による蛋白質構造変化の解析" 生物物理. 37. 225-226 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 七田芳則: "生物物理から見た生命像4 視る感覚 「光のセンサー;視物質」(分担執筆)" 吉岡書店, 127 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 七田芳則: "視覚、図で学び絵で見るレーザー科学入門(分担執筆)" 丸善, (CD-ROM) (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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