Target molecule recognition and redox regulation by the chloroplast thioredoxin

• Project/Area Number: 17370015
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 植物生理・分子
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: HISABORI Toru Tokyo Institute of Technology, Chemical Resources Laboratory, Associate Professor (40181094)
• Co-Investigator(Kenkyū-buntansha): NUREKI Osamu Tokyo Institute of Technology, Department of Biological Information, Graduate School of Bioscience and Biotechnology, Professor (10272460)
• Project Period (FY): 2005 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥15,440,000 (Direct Cost: ¥14,600,000、Indirect Cost: ¥840,000); Fiscal Year 2007: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2006: ¥4,100,000 (Direct Cost: ¥4,100,000); Fiscal Year 2005: ¥7,700,000 (Direct Cost: ¥7,700,000)
• Keywords: thioredoxin / redox regulation / disulfide bond / protein-protein interaction / cyanobacteria / リンゴ酸脱水素酵素 / 標的酵素 / 1分子観察 / 蛋白質間相互作用

Research Abstract

A. Interaction between thioredoxins and their targets
We first studied the interaction between the plant cytosolic thioredoxin (Trx-hl) and the cytosolic malate dehydrogenase and successfully revealed that this malate dehydrogenase is the newly identified Trx-target protein. We then studied the counterpart proteins of thioredoxins from photosynthetic bacteria and cyanobacteria to reveal the multiplicity of the role of thioredoxins. In addition, we clarified the difference of the roles of two redox-equivalent supply pathways, ferredoxin-dependent thioredoxin reductase pathway and NADPH dependent thioredoxin reductase pathway in the cyanobacteria Synechocystis sp. PCC6803.
B. Structural analysis of the protein-protein interaction between thioredoxins and their targets.
One of the most interesting questions are why thioredoxins can recognize various proteins, which contains the redox sensitive disulfide bond on the molecular surface, as their target. In order to answer this query, we tried to solve the crystal structures of the thioredoxin-target protein cocomplex using X-ray crystallography. Although we have tried to efficiently obtain the co-complexexes under the various chemical conditions, we failed to set up the suitable experimental conditions to obtain the complex proteins during the research term and could not solve any structures which should be important to know the amino acid residues which may involve into the interaction.

Research Products

• [Journal Article] Binary reducing equivalent pathways using NADPH-thioredoxin reductase and ferredoxin-thioredoxin reductase in the cyanobacterium Synechocystis sp. strain PCC 6803. (2008)
  • Author(s): Hishiya S, Hatakeyama W, Mizota Y, Hosoya-Matsuda N, Motohashi K, Ikeuchi M, Hisabori T.
  • Journal Title: Plant Cell Phyisology 49 Pages: 11-18
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Binary reducing equivalent pathways using NADPH-thioredoxin reductase and ferredoxin-thioredoxin reductase in the cyanobacterium Synechocystis sp. strain PCC 6803. (2008)
  • Author(s): S, Hishiya, W, Hatakeyama, Y, Mizota, N, Hosoya-Matsuda, K, Motohashi, M, Ikeuchi, T., Hisabori
  • Journal Title: Plant Cell Phyisology 49 Pages: 49-49
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Binary reducing equivalent pathways using NADPH-thioredoxin reductase and ferredoxin-thioredoxin reductase in the cyanobacterium Synechocystis sp. strain PCC 6803. (2008)
  • Author(s): Hishiya, S. et. al.
  • Journal Title: Plant Cell Phyisology 49 Pages: 11-18
  • Peer Reviewed
• [Journal Article] Towards a Functional Dissection of Thioredoxin Networks in Plant Cells. (2007)
  • Author(s): Hisabori T, Motohashi K, Hosoya-Matsuda N, Ueoka-Nakanishi H, Romano PG.
  • Journal Title: Photochem Photobiol. 83 Pages: 145-151
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The significance of type II and PrxQ Peroxiredoxins for antioxidative stress response in the purple bacterium Rhodobacter sphaeroides. (2007)
  • Author(s): Wakita M, Masuda S, Motohashi K, Hisabori T, Ohta H, Takamiya K.
  • Journal Title: J. Biol. Chem. 282 Pages: 27792-27801
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The CHLI1 subunit of Arabidopsis thaliana magnesium chelatase is a target protein of the chloroplast thioredoxin. (2007)
  • Author(s): Ikegami A, Yoshimura N, Motohashi K, Takahashi S, Romano PG, Hisabori T, Takamiya K, Masuda T.
  • Journal Title: J. Biol. Chem. 282 Pages: 19282-19291
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Towards a Functional Dissection of Thioredoxin Networks in Plant Cells (2007)
  • Author(s): T., Hisabori, K, Motohashi, N, Hosoya-Matsuda, H., Ueoka-Nakanishi, PG., Romano
  • Journal Title: Photochem Photobiol 83 Pages: 145-151
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The significance of type II and PrxQ peroxiredoxins for antioxidative stress response in the purple bacterium Rhodobacter sphaeroides (2007)
  • Author(s): M, Wakita, S, Masuda, K, Motohashi, T, Hisabori, H, Ohta, K., Takamiya
  • Journal Title: J. Biol. Chem 282 Pages: 27792-27801
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The CHLI1 subunit of Arabidopsis thaliana magnesium chelatase is a target protein of the chloroplast thioredoxin (2007)
  • Author(s): A, Ikegami, N, Yoshimura K, Motohashi, S, Takahashi, PG, Romano, T, Hisabori, K, Takamiya, T., Masuda
  • Journal Title: J. Biol. Chem 282 Pages: 19282-19291
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The significance of type II and PrxQ peroxiredoxins for antioxidative stress response in the purple bacterium Rhodobacter sphaeroides. (2007)
  • Author(s): Wakita M. et. al.
  • Journal Title: J. Biol. Chem. 282 Pages: 27792-27801
  • Peer Reviewed
• [Journal Article] The CHLI1 subunit of Arabidopsis thaliana magnesium chelatase is a target protein of the chloroplast thioredoxin. (2007)
  • Author(s): Ikegami, A. et. al.
  • Journal Title: J. Biol. Chem. 282 Pages: 19282-19291
  • Peer Reviewed
• [Journal Article] Towards a Functional dissection of Thioredoxin Networks in Plant Cells. (2007)
  • Author(s): Hisabori, T., et al.
  • Journal Title: Photochem Photobiol. 83 Pages: 145-151
• [Journal Article] Thioredoxin-h1 reduces and reactivates the oxidized cytosolic malate dehydrogenase dimer in higher plants (2006)
  • Author(s): Hara S, Motohashi K, Arisaka F, Romano PG, Hosoya-Matsuda N, Kikuchi N, Fusada N, Hisabori T.
  • Journal Title: J. Biol. Chem. 281(43) Pages: 32065-32071
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Thioredoxin-hl reduces and reactivates the oxidized cytosolic malate dehydrogenase dimer in higher plants (2006)
  • Author(s): Hara, s., et al.
  • Journal Title: J. Biol. Chem. 281(43) Pages: 32065-32071
• [Journal Article] Thioredoxin affinity chromatography: a useful method for further understanding the thioredoxin network (2005)
  • Author(s): Hisabori, T., Hara, S., Fujii, T., Yamazaki, D., Hosoya-Matusda, N., Motohashi, K.
  • Journal Title: Journal of Experimental Botany 56(416) Pages: 1463-1468
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Thioredoxin affinity chromatography : a useful method for further understanding the thioredoxin network (2005)
  • Author(s): T., Hisabori, S. Hara, T., Fujii, D., Yamazaki, N., Hosoya-Matusda, K., Motohashi
  • Journal Title: Journal of Experimental Botany 56 Pages: 1463-1468
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Thioredoxin-hl reduces and reactivates the oxidized cytosolic malate dehydrogenase dimer in higher plants (2005)
  • Author(s): S., Hara, K., Motohashi, F., Arisaka, PG., Romano, N., Hosoya-Matsuda, N., Kikuchi, N., Fusada, T., Hisabori
  • Journal Title: J. Biol. Chem 281 Pages: 32065-32071
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Thioredoxin affinity chromatography : a useful method for further understanding the thioredoxin network (2005)
  • Author(s): Hisabori, T., Hara, S., Fujii, T., Yamazaki, D., Hosoya-Matusda, N., Motohashi, K.
  • Journal Title: Journal of Experimental Botany 56(416) Pages: 1463-1468
• [Presentation] 細胞質型フルクトースビスリン酸アルドラーゼの酸化還元制御機構 (2008)
  • Author(s): 丸井弘嗣, 原怜, 本橋健, 久堀徹
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Anabaena sp. PCC 7120におけるチオレドキシン還元標的タンパク質の探査 (2008)
  • Author(s): 前田真希、松田直美、久堀徹、井上和仁
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 緑色硫黄細菌Chlorobaculum tepidum TLSのチオレドキシンによるリンゴ酸デヒドロゲナーゼの活性調節 (2008)
  • Author(s): 松田直美、高橋昭博、久堀徹、井上和仁
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] チオレドキシンはカルシウム依存キナーゼをレドックス調節する (2008)
  • Author(s): 中西華代、佐塚隆志、森仁志、久堀徹
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ラン藻における翻訳伸長因子EF-Gの酸化還元と翻訳制御 (2008)
  • Author(s): 小島浩治、大下将、久堀徹、林秀則、西山佳孝
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Redox regulation of the cytosolic fructose 1,6-bis phosphate aldolase (2008)
  • Author(s): K., Marui, S., Hara, K., Motohashi, T., Hisabori
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Potential target proteins of thioredoxin in cyanobacterium Anabaena sp. PCC 7120 (2008)
  • Author(s): M., Maeda, N., Matsuda, T., Hisabori, K., Inoue
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Thioredoxin regulate malate dehydrogenase in green sulfur bacterium Chlorobaculum tepidum TLS (2008)
  • Author(s): N., Matsuda, A., Takahashi, T, Hisabori, K., Inoue
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Redox regulation of Ca-dependent protein kinase by thioredoxin (2008)
  • Author(s): H., Nakanishi, T, Sazuka, H., Mori, T., Hisabori
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Regulation of translation caused by the redox change of EF-G of cyanobacteria (2008)
  • Author(s): K., Kojima, S., Ohshita, T., Hisabori, H., Hayashi, Y., Nishiyama
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-21
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] シロイヌナズナのMg-キラターゼのサブユニットCHLIアイソフォームの機能解析 (2008)
  • Author(s): 小林康一、望月伸悦、吉村奈穂、本橋健、久堀徹、増田建
  • Organizer: 日本植物生理学会第49回年会
  • Place of Presentation: 札幌コンベンションセンター
  • Year and Date: 2008-03-19
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Functional analysis of isoforms of CHLI subunit of Mg-chelatase in Arabidopsis thaliana (2008)
  • Author(s): K., Kobayashi, N., Mochizuki, N., Yoshimura, K., Motohashi, T., Hisabori, T., Masuda
  • Organizer: 49th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Sapporo
  • Year and Date: 2008-03-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 遺伝子破壊によるSynechocystis sp. PCC 6803の二種類のチオレドキシン還元経路の解析 (2007)
  • Author(s): 畠山和佳子、菱谷 昇子, 松田 直美, 本橋 健, 池内 昌彦, 久堀 徹
  • Organizer: ラン藻研究会
  • Place of Presentation: かずさアーク
  • Year and Date: 2007-12-03
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Study on the two thioredoxin reduction pathways in Synechocystis sp. PCC6803 by gene disruption. (2007)
  • Author(s): W., Hatakeyama, S., Hishiya, N., Matsuda, K., Motohashi, M., Ikeuchi, T., Hisabori
  • Organizer: Meeting for cyanobacteria
  • Place of Presentation: Kazusa, Chiba
  • Year and Date: 2007-12-03
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Synechocystis sp. PCC6803の二種類のチオレドキシン還元経路 (2007)
  • Author(s): 菱谷 昇子, 溝田 陽子, 松田 直美, 本橋 健, 池内 昌彦, 久堀 徹
  • Organizer: 日本植物学会第71回大会
  • Place of Presentation: 東京理科大学野田キャンパス
  • Year and Date: 2007-09-09
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Two pathways for thioredoxin reduction in Synechocystis sp. PCC6803 (2007)
  • Author(s): S., Hishiya, Y., Mizota, N., Matsuda, K., Motohashi, M., Ikeuchi, T., Hisabori
  • Organizer: 71th Annual Meeting of the Botanical Society Japan
  • Place of Presentation: Tokyo Univ. Sci
  • Year and Date: 2007-09-09
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Synechocystis sp. PCC6803の二種類のチオレドキシン還元経路 (2007)
  • Author(s): 久堀 徹
  • Organizer: 日本植物学会
  • Place of Presentation: 東京理科大学野田キャンパス
  • Year and Date: 2007-09-09
• [Presentation] タンパク質合成系の酸化ストレス障害とレドックス制御 (2007)
  • Author(s): 西山佳孝、小島幸治、大下将、久堀徹、林秀則
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] チオレドキシン標的候補として細胞膜アンカー型信号伝達分子を同定した (2007)
  • Author(s): 中西華代、佐塚隆志、前島正義、久堀徹
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 紅色細菌Rhodobacter sphaeroidesにおけるペルオキシレドキシンの生理機能解析 (2007)
  • Author(s): 脇田将裕、増田真二、本橋健、久堀徹、太田啓之、高宮建一郎
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] シロイヌナズナのMg-キラターゼのサブユニットCHLI1におけるチオレドキシン標的システイン残基の同定 (2007)
  • Author(s): 吉村菜穂、池上陽紀、本橋健、高橋重一、久堀徹、増田建
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] ラン藻の翻訳伸長因子EF-Gとチオレドキシンの相互作用 (2007)
  • Author(s): 大下将、小島幸治、久堀徹、西山佳孝、林秀則
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 緑色硫黄細菌Chlorobium tepidum TLSのチオレドキシンシステム (2007)
  • Author(s): 松田直美、高橋昭博、伊里耕太、久堀徹、井上和仁
  • Organizer: 第48回日本植物生理学会年会
  • Place of Presentation: 愛媛大学
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Oxidative stress and redox regulation of protein synthesis (2007)
  • Author(s): Y., Nishiyama, K., Kojima, S., Oshita, T., Hisabori, H., Hayashi
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Possible thioredoxin targets of signaling molecules in plasma membranes (2007)
  • Author(s): H., Nakanishi, T., Sazuka, M., Maeshima, T., Hisabori
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Study of peroxiredoxin of Rhodobacter sphaeroides (2007)
  • Author(s): M., Wakita, S., Masuda, K., Motohashi, T., Hisabori, H., Ohta, K-I., Takamiya
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Identification of cysteine residues involved in thioredoxin-dependent redox regulation of CHLI1, a subunit of Mg-chelatase in Arabidopsis thaliana (2007)
  • Author(s): N., Yoshimura, A., Ikegami, K., Motohashi, J., Takahashi, T., Hisabori, T., Masuda
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Interaction between thioredoxin and EF-G of cyanobacteria (2007)
  • Author(s): S., Ohshita, K., Kojima, T., Hisabori, Y., Nishiyama, H., Hayashi
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Thioredoxin system of the Green suffer bacteria Chlorobium tepidum TLS (2007)
  • Author(s): N., Matsuda, A., Takahashi, K., Isato, T., Hisabori, K., Inoue
  • Organizer: 48th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Ehime Univ
  • Year and Date: 2007-03-28
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 高等植物細胞質のリンゴ酸脱水素酵素は新規のチオール酵素である (2006)
  • Author(s): 原怜, 本橋健, 久堀徹
  • Organizer: 第47回日本植物生理学会年会
  • Place of Presentation: 筑波大学
  • Year and Date: 2006-03-19
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] The plant cytosolic malate dehydrogenase is the newly identified thiol enzyme (2006)
  • Author(s): S., Hara, K., Motohashi, T., Hisabori
  • Organizer: 47th Annual Meeting of the Japanese Society of Plant Physiologist
  • Place of Presentation: Tsukuba Univ
  • Year and Date: 2006-03-19
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 高等植物細胞質型リンゴ酸脱水素酵素の酸化還元調節 (2005)
  • Author(s): 原怜, 本橋健, 久堀徹
  • Organizer: 第78回日本生化学会大会
  • Place of Presentation: 神戸国際展示場
  • Year and Date: 2005-10-20
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Redox regulation of the plant cytosolic malate dehydrogenase (2005)
  • Author(s): S., Hara, K., Motohashi, T., Hisabori
  • Organizer: 78th Japan Biochmical Society Meeting
  • Place of Presentation: Kobe
  • Year and Date: 2005-10-20
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] シアノバクテリアチオレドキシンの相互干渉とレドックス制御機能 (2005)
  • Author(s): 溝田 陽子, 松田 直美, 本橋 健, 久堀 徹
  • Organizer: 日本植物学会第69回大会
  • Place of Presentation: 富山大学
  • Year and Date: 2005-09-06
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Interferences of the cyanobacterial thioredoxins and their redox regulation (2005)
  • Author(s): Y., Mizota, N., Matsuda, K., Motohashi, T., Hisabori
  • Organizer: 69th Annual Meeting of the Botanical Society Japan
  • Place of Presentation: Toyama Univ
  • Year and Date: 2005-09-06
  • Description: 「研究成果報告書概要(欧文)」より