Development of photo-driven motor proteins using phtochromic molecules

• Project/Area Number: 14580677
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biophysics
• Research Institution: Soka University
• Principal Investigator: MARUTA Shinsaku Soka University, Dept.of Bioinformatics, Professor, 工学部, 教授 (40231732)
• Co-Investigator(Kenkyū-buntansha): 茶圓 茂 日本大学, 理工学部, 教授 (60142452)
• Project Period (FY): 2002 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000); Fiscal Year 2002: ¥1,100,000 (Direct Cost: ¥1,100,000)
• Keywords: molecular motor / myosin / kinesin / photochromic molecule / azobenzene / laser / energy transduction / ATPase / 化学修飾 / X線溶液散乱 / 蛍光 / 構造変化

Research Abstract

ATP driven-motor proteins transduce chemical energy generated from ATP hydrolysis into the physical motion energy with the mechanical system like cam-shaft. It is expected that introduction of artificial energy input device into the energy transducing site of motor protein, enable the driving or regulating of motor proteins without ATP. In the present study, photochromic molecules which change their conformation reversibly with high response due to the isomerization by ultraviolet (UV) and visible light (VIS) irradiation, have been utilized as energy input device. Especially azobenzene and spiropyran derivatives, which change their characteristics dramatically on isomerization by UV-VIS light irradiation, might work effectively.
In this study, several kinds of azobenzene and spiropyran derivatives, which can bind to cysteine side chain covalently, have been synthesized. And several motor protein mutants, which have one reactive cysteine at the specific site, have been also prepared. It has been demonstrated that for skeletal muscle myosin motor domain, the localized conformational changes resulting from isomerization on azobenzene derivative cross-linking of reactive cysteine residues, SH1 and SH2 induced swing in the lever arm portion. It was shown that the incorporated photochromic group acts as a mechanical energy input in a motor protein. The new technique using photochromic molecules may be applicable to other energy transducing and regulatory proteins.

Research Products

• [Journal Article] Preparation and characterization of a novel rice plant specific kinesin (2006)
  • Author(s): Umeki, N., Mitsui, T., Umezu, N., kondo, K., Maruta, S.
  • Journal Title: Journal of Biochemistry 139 Pages: 645-654
  • NAID: 10018845372
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Intermolecular cross-linking of a novel rice kinesin K16 motor domain with a photoreactive ATP derivative (2006)
  • Author(s): Umeki, N., Mitsui, T., Koike, Y., Maruta, S.
  • Journal Title: Journal of Biochemistry 139 Pages: 831-836
  • NAID: 10018844848
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Conformational change of the loop L5 in rice kinesin motor domain induced by nucleotide binding (2006)
  • Author(s): Umeki, N., Mitsui, T., Kondo, K., Maruta, S.
  • Journal Title: Journal of Biochemistry 139 Pages: 857-864
  • NAID: 10018844929
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Preparation and characterization of a novel rice plant specific kinesin (2006)
  • Author(s): Umeki, N., Mitsui, T., Umezu, N., Kondo, K., Maruta, S
  • Journal Title: J.Biochem. 139 Pages: 645-654
  • NAID: 10018845372
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Intermolecular cross-linking of a novel rice kinesin K16 motor domain with a photoreactive ATP derivative (2006)
  • Author(s): Umeki, N., Mitsui, T., Koike, Y., Maruta, S.
  • Journal Title: J.Biochem. 139 Pages: 831-836
  • NAID: 10018844848
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Conformational change of the loop L5 in rice kinesin motor domain induced by nucleotide binding (2006)
  • Author(s): Umeki, N., Mitsui, T., Kondo, K., Maruta, S.
  • Journal Title: J.Biochem. 139 Pages: 857-864
  • NAID: 10018844929
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Preparation and characterization of a novel rice plant specific kinesin (2006)
  • Author(s): Umeki, N., Mitsui, T., Umezu, N., Kondo, K., Maruta, S.
  • Journal Title: Journal of Biochemistry (In press)
  • NAID: 10018845372
• [Journal Article] Intermolecular cross-linking of a novel rice kinesin K16 motor domain with a photoreactive ATP derivative (2006)
  • Author(s): Umeki, N., Mitsui, T., Koike, Y., Maruta, S.
  • Journal Title: Journal of Biochemistry (In press)
  • NAID: 10018844848
• [Journal Article] Conformational change of the loop L5 in rice kinesin motor domain induced by nucleotide binding (2006)
  • Author(s): Umeki, N., Mitsui, T., Kondo, K., Maruta, S.
  • Journal Title: Journal of Biochemistry (In press)
  • NAID: 10018844929
• [Journal Article] Three-dimensional structural analysis of individual myosin heads undervarious functional states (2004)
  • Author(s): Katayama, E., Ichise, N., Yaeguchi, N., Yoshizawa, T., Maruta, S., Baba, N.
  • Journal Title: Adv. In Exp. Med. And Biol. 538 Pages: 295-304
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Interaction of myosin-ADP-fluorometal complexes with fluorescent probes and direct observation using quick-freeze deep-etch electron microscopy (2004)
  • Author(s): Maruta, S., Uyehara, Y., Aihara, T., Katayama, E.
  • Journal Title: Journal of Biochemistry 136 Pages: 57-64
  • NAID: 10016200785
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Incorporation of an azobenzene derivative into the energy transducing site of skeletal muscle myosin results in photo-induced conformational changes (2004)
  • Author(s): Umeki, N., Yoshizawa, T., Sugimoto, Y., Mitsui, T., Wakabayashi, K., Maruta, S.
  • Journal Title: Journal of Biochemistry 136 Pages: 839-846
  • NAID: 10016204499
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Three-dimensional structural analysis of individual myosin heads undervarious functional states (2004)
  • Author(s): Katayama, E., Ichise, N., Yaeguchi, N., Yoshizawa, T., Maruta, S., Baba, N.
  • Journal Title: Adv.In Exp.Med.And Biol. 538 Pages: 295-304
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Interaction of myosin ・ADP・ fluorometal complexes with fluorescent probes and direct obervation using quick-freeze deep-etch electron microscopy (2004)
  • Author(s): Maruta, S., Uyehara, Y., Aihara, T., Katayama, E.
  • Journal Title: J.Biochem. 136 Pages: 57-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Incorporation of an azobenzene derivative into the energy transducing site of skeletal muscle myosin results in photo-induced conformational changes (2004)
  • Author(s): Umeki, N., Yoshizawa, T., Sugimoto, Y., Mitsui, T., Wakabayashi, K., Maruta, S.
  • Journal Title: J.Biochem. 136 Pages: 839-846
  • NAID: 10016204499
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Three-dimensional structural analysis of individual myosin heads undervarious functional states. (2004)
  • Author(s): Katayama, E., Ichise, N., Yaeguchi, N., Yoshizawa, T., _Maruta, S., Baba, N.
  • Journal Title: Adv.In Exp.Med. And Biol. 588 Pages: 295-304
• [Journal Article] Interaction of myosin・ADP・fluorometal complexes with fluorescent probes and direct observation using quick-freeze deep-etch electron microscopy (2004)
  • Author(s): Maruta, S., Uyehara, Y., Aihara, T., Katayama, E.
  • Journal Title: Journal of Biochemistry 136 Pages: 57-64
  • NAID: 10016200785
• [Journal Article] Incorporation of an azobenzene derivative into the energy transducing site of skeletal muscle myosin results in phto-induced conformational change (2004)
  • Author(s): Umeki, N., Yoshizawa, T., Sugimoto, Y., Mitsui, T., Wakabayashi, K., Maruta
  • Journal Title: Journal of Biochemistry 136 Pages: 839-846
• [Publications] Komaba, S., Inoue, A., Maruta, S., Hosoya, H., Ikebe, _M.: "Determination of Human Myosin III as a Motor Protein Having a Protein Kinase Activity"J.Biol.Chem.. 278. 21352-21360 (2003)
• [Publications] E.Katayama, N.Ichise, N.Yaeguchi, T.Yoshizawa, S.Maruta, N.Baba: "Three-dimensional structural analysis of individual myosin heads undervarious functional states"Adv.in Exp.Med. and Biol.. 538. 295-304 (2004)
• [Publications] Maruta, S., Mizukura, Y., Chaen, S.: "Interaction of a New Fluorescent ATP Analogue with Skeletal Muscle Myosin Subfragment-1"J. Biochem.. 131. 905-911 (2002)
• [Publications] Maruta, S., Mizukura, Y.: "Analysis of the Conformational Change of Myosin during ATP Hydrolysis Using Fluorescence Resonance Energy Transfer"J. Biochem.. 132. 471-482 (2002)
• [Publications] Shibuya, H., Kondo, K., Kimura, N., Maruta, S.: "Formation and Characterization of Kinesin・ADP・Fluorometal Complexes"J. Biochem.. 132. 573-579 (2002)