Synthesis of functional probes toward elucidation of the function of respiratory enzymes of Caenorhabditis elegans.

• Project/Area Number: 15380083
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bioproduction chemistry/Bioorganic chemistry
• Research Institution: KYOTO UNIVERSITY
• Principal Investigator: MIYOSHI Hideto Kyoto University, Agriculture, Associate Professor, 農学研究科, 助教授 (20190829)
• Co-Investigator(Kenkyū-buntansha): KITA Kiyoshi University of Tokyo, Medicine, Professor, 大学院・医学系研究科, 教授 (90134444)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥13,700,000 (Direct Cost: ¥13,700,000); Fiscal Year 2004: ¥5,300,000 (Direct Cost: ¥5,300,000); Fiscal Year 2003: ¥8,400,000 (Direct Cost: ¥8,400,000)
• Keywords: Respiratory Inhibitor / Respiratory Enzymes / Mitochondria / アセトゲニン / 構造活性相関

Research Abstract

To identify the binding subunit of fenpyroximate, a very potent inhibitor of bovine heart mitochondrial complex I, we synthesized ^3H-labeled photolabile fenpyroximate ([^3H](trifluoromethyl)-phenyldiazirinyl fenpyroximate, TDF). TDF appeared to elicit very potent inhibition of bovine heart mitochondrial complex I being nearly equivalent to that of fenpyroximate.
On the basis of photoaffinity studies with bovine heart complex I in collaboration with Dr.Takao Yagi (The Scripps Research Institute, USA), the binding site of TDF was shown to be subunit ND5 which is thought to exist in membrane arm of the enzyme. Since antibody to the C-terminal region of the ND5 subunit only weakly recognized the subunit, we identified the ND5 by 2D-electrophoresis (SDS-PAGE). Our result is the first study wherein the subunit located in membrane arm was identified as the inhibitor binding subunit. Our study strongly suggests that subunit(s) located in membrane arm may be involved in the proton-pumping activity of complex I.

Research Products

• [Journal Article] Synthesis and inhibition mechanism of Dlac-acetogenins : a novel type of inhibitor of bovine heart mitochondrial complex I. (2005)
  • Author(s): Ichimaru, N., Murai, M., Abe, M., Hamada, T., Yamada, Y., Makino, S., Nishioka, T., Makabe, H., Makino, A., Kobayashi, T., Miyoshi, H.
  • Journal Title: Biochemistry 44 Pages: 816-825
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis of two possible diastereomers of reticulatain-I. (2004)
  • Author(s): Makabe, H. et al.
  • Journal Title: Tetrahedron Lett. 45 Pages: 973-977
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Rhodoquinone Reaction Site of Mitochondrial Complex I in Parasitic Helminth. (2004)
  • Author(s): Yamashita, T., Ino, T., Miyoshi, H., Sakamoto, K., Osanai, A., Nakamaru-Ogiso, E., Kita, K.
  • Journal Title: Ascaris suum.Biochim.Biophys.Acta (Bioenergetics) 1608 Pages: 97-103
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis of two possible diastereomers of reticulatain-1 (2004)
  • Author(s): Makabe, H., Miyawaki, A., Takahashi, R., Hattori, Y., Konno, H., Abe, M., Miyoshi, H.
  • Journal Title: Tetrahedron Lett. 45 Pages: 973-977
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Occurrence of a bound ubiquinone and its function in Escherichia coli membrane-bound quinoprotein glucose dehydrogenase. (2004)
  • Author(s): Elias, M.D., Nakamura, S., Migita, C.T., Miyoshi, H., Toyama, H., Matsushita, K., Adachi, O., Yamada, M.
  • Journal Title: J.Biol.Chem. 279 Pages: 3078-3083
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis and mitochondrial complex I inhibition of dihydroxy-cohibin A, non-THF annonaceous acetogenin analogue (2004)
  • Author(s): Konno, H., Hiura N., Makabe, H., Abe, M., Miyoshi, H.
  • Journal Title: Bioorg.Med.Chem.Lett. 14 Pages: 629-632
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Essential structural features of acetogenins : role of hydroxy groups adjacent to the bis-THF rings. (2004)
  • Author(s): Abe, M., Kenmochi, A., Ichimaru, N., Hamada, T., Nishioka, T., Miyoshi, H.
  • Journal Title: Bioorg.Med.Chem.Lett. 14 Pages: 779-782
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis and Inhibitory Action of Novel Acetogenin Mimics with Bovine Heart Mitochondrial Complex I. (2004)
  • Author(s): Hamada, T., Ichimaru, N., Abe, M., Fujita, D., Kenmochi, A., Nishioka, T., Zwicker, K., Brandt, U., Miyoshi, H.
  • Journal Title: Biochemistry 43 Pages: 3651-3658
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Direct evidence for the interaction of stigmatellin with a protonated acidic group in the bc_1 complex from S.cerevisiae as monitored by FTIR difference spectroscopy and ^<13>C specific labeling. (2004)
  • Author(s): Ritter, M., Palsdottir, H., Abe, M., Mantele, W., Hunte, C., Miyoshi, H., Hellwig, P.
  • Journal Title: Biochemistry 43 Pages: 8439-8446
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Total synthesis of cis-solamin and its inhibitory action with bovine heart mitochondrial complex I. (2004)
  • Author(s): Makabe, H., Hattori, Y., Kimura, Y., Konno, H., Abe, M., Miyoshi, H., Tanaka, A., Oritani, T.
  • Journal Title: Tetrahedron 60 Pages: 10651-10657
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis of two possible diasteromers of reticulatain-I. (2004)
  • Author(s): Makabe, H. et al.
  • Journal Title: Tetrahedron Lett. 45 Pages: 973-977
• [Journal Article] The ND5 subunit was labeled by a photoaffinity analogue of fenpyroximate in bovine mitochondria complex I. (2003)
  • Author(s): Nakamura-Ogiso, E. et al.
  • Journal Title: Biochemistry 42 Pages: 746-754
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Architecture of succinate dehydrogenase and reactive oxygen species generation (2003)
  • Author(s): Yankovskaya, V. et al.
  • Journal Title: Science 299 Pages: 700-704
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Atpenins, potent and specific inhibitors of mitochondrial complex II (succinate-ubiquinone oxidoredutase). (2003)
  • Author(s): Miyadera, H. et al.
  • Journal Title: Proc.Natl.Acad.Sci.USA 100 Pages: 473-477
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Asymmetric binding of the high-affinity Q_H ubisemiquinone in quinol oxidase (bo_3) from Escherichia coli by mulifrequency electron paramagnetic resonance spectroscopy. (2003)
  • Author(s): Grimaldi S. et al.
  • Journal Title: Biochemistry 42 Pages: 5632-5639
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Synthesis and Inhibitory Action of Novel Acetogenin Mimics with Bovine Heart Mitochondrial Complex I. (2003)
  • Author(s): Hamada, T. et al.
  • Journal Title: Biochemistry 43 Pages: 3651-3658
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The ND5 subunit was labeled by a photoaffinity analogue of fenpyroximate in bovine mitochondrial complex I. (2003)
  • Author(s): Nakamaru-Ogiso, E., Sakamoto, K., Matsuno-Yagi, A., Miyoshi, H., Yagi, T.
  • Journal Title: Biochemistry 42 Pages: 746-754
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Architecture of succinate dehydrogenase and reactive oxygen species generation (2003)
  • Author(s): Yankovskaya, V., Horsefield, R., Tornroth, S., Luna-Chavez, C., Miyoshi, H., Leger, C., Byrne, B., Cecchini, G., Iwata, S.
  • Journal Title: Science 299 Pages: 700-704
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Atpenins, potent and specific inhibitors of mitochondrial complex II (succinate-ubiquinone oxidoreductase). (2003)
  • Author(s): Miyadera, H., Shiomi, K., Ui, H., Yamaguchi Y., Masuma, R., Tomoda, H., Miyoshi, H., Osanai, A., Kita, K., Ohmura, S.
  • Journal Title: Proc.Natl.Acad.Sci.USA 100 Pages: 473-477
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Asymmetric binding of the high-affinity Q_H ubisemiquinone in quinol oxidase (bo_3) from Escherichia coli studied by multifrequency electron paramagnetic resonance spectroscopy (2003)
  • Author(s): Grimaldi S., Ostermann, T., Weiden, N., Mogi, T., Miyoshi, H., Lidwig, B., Michel, H., Prinsner, T.F., MacMillan, F.
  • Journal Title: Biochemistry 42 Pages: 5632-5639
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Complex II from phototrophic purple bacterium Rhodoferax fermentans displays rhodoquinol-fumarate reductase activity. (2003)
  • Author(s): Miyadera, H., Hiraishi, A., Miyoshi, H., Sakamoto, K., Mineki, R., Murayama, K., Nagashima, K., Matsuura, K., Kojima, S., Kita, K.
  • Journal Title: Eur.J.Biochem. 270 Pages: 1863-1874
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Isolation and characterization of the stage-specific cytochrome b small subunit (CybS) of Ascaris suum complex II from the aerobic respiratory chain of larval mitochondria. (2003)
  • Author(s): Amino, H., Osanai, A., Miyadera, H., Shinjyo, N., Tomitsuka, E., Taka, H.Mineki, R.Murayama, K.Takamiya, S.Aoki, T.Miyoshi, H., Sakamoto, K Kojima, S., Kita, K.
  • Journal Title: Mol.Biochem.Parasitol. 128 Pages: 175-186
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Synthesis and Inhibitory Activity of Ubiquinone-Acetogenin Hybrid Inhibitor with Bovine Mitochondrial Complex I. (2003)
  • Author(s): Yabunaka, H., Abe, M., Kenmochi, A., Hamada, T., Nishioka T., Miyoshi, H.
  • Journal Title: Bioorg.Med.Chem.Lett. 13 Pages: 2385-2388
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Characterization of inhibitor binding sites of mitochondrial complex I using fluorescent inhibitor (2003)
  • Author(s): Ino, T., Nishioka, T., Miyoshi, H.
  • Journal Title: Biochim.Biophys.Acta (Bioenergetics) 1605 Pages: 15-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] 山下哲夫: "Rhodoquinone reaction site of mitochondrial complex I in parasitic helminth, Ascaris sum"Biochim.Biophys.Acta. 1608. 97-103 (2004)
• [Publications] 真壁秀文: "Synthesis of two possible diastereomers of reticulatiain-1"Tetrahedron Lett.. 45. 973-977 (2004)
• [Publications] M.D.Elias: "Occurrence of a bound ubiquinone and its function in Escherichia coli membrane-bound quinoprotein glucose dehydrogenase"J.Biol.Chem.. 279. 3078-3083 (2004)
• [Publications] 安部真人: "Essential structural features of acetogenins : role of hydroxy groups adjacent to the bis-THF ring"Bioorg.Med.Chem.Lett.. 14. 779-782 (2004)