Development of the expression system of helminthic mitochondrial quinol-fumarate reductase in bacteria.

• Project/Area Number: 23590484
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Parasitology (including Sanitary zoology)
• Research Institution: Hirosaki University
• Principal Investigator: SAKAMOTO Kimitoshi 弘前大学, 農学生命科学部, 准教授 (50361465)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 蠕虫 / ミトコンドリア呼吸鎖

Research Abstract

Quinol-fumarate reductase (QFR) is the key enzyme for the anaerobic respirtory chain of hypoxia adapted helminthes. QFR is characteristic by its effective catalytic activity for the reverse reaction of aerobic respiratory complex II. In the aim of elucidating the mechanisms of enhancing reverse reaction, the development of expression system in bacteria was planned. At first, all the genes for the subunits of QFR from Echinococcus multilocularis were cloned. N terminal amino acid of them were also determined from the QFR in mitochondria. Complex II gene from the expression host, Rhodospirillum rubrum was knocked out. Finally, plasmid for expressing Echonococcus QFR was prepared.

Research Products

• [Journal Article] Pharmacophore identification of ascofuranone, potent inhibitor of cyanide-insensitive alternative oxidase of Trypanosoma brucei (2013)
  • Author(s): Saimoto H, Kido Y, Haga Y, Sakamoto K, Kita K
  • Journal Title: The Journal of Biochemistry Volume: 153 Pages: 267-273
• [Journal Article] Structure of the trypanosome cyanide-insensitive alternative oxidase (2013)
  • Author(s): Shiba T, Kido Y, Sakamoto K, Inaoka DK, Tsuge C, Tatsumi R, Takahashi G, Balogun EO, Nara T, Aoki T, Honma T, Tanaka A, Inoue M, Matsuoka S, Saimoto H, Moore AL, Harada S, Kita K
  • Journal Title: Proceedings of the National Academy of Sciences of the United States of America Volume: 110 Pages: 4580-4585
• [Journal Article] Cloning and characterization of hypoxia-inducible factor-1 subunits from Ascaris suum - a parasitic nematode highly adapted to changes of oxygen conditions during its life cycle (2013)
  • Author(s): Goto, M., Amino, H., Nakajima, M., Tsuji, N., Sakamoto, K., Kita, K.
  • Journal Title: Gene Volume: 516 Issue: 1 Pages: 39-47
  • DOI: 10.1016/j.gene.2012.12.025
  • Peer Reviewed
• [Journal Article] Age-related changes in the activities of respiratory chain complexes and mitochondrial morphology in Drosophila (2012)
  • Author(s): Oda Y, Yui R, Sakamoto K, Kita K, Matsuura ET
  • Journal Title: Mitochondrion Volume: 12 Pages: 345-351
• [Journal Article] Crystal structure of mitochondrial quinol-fumarate reductase from the parasitic nematode Ascaris suum (2012)
  • Author(s): Shimizu H, Osanai A, Sakamoto K, Inaoka D K, Shiba T, Harada S, Kita K
  • Journal Title: The Journal of Biochemistry Volume: 151 Pages: 589-592
  • NAID: 10031166100
• [Journal Article] Ukulactones A and B, new NADH-fumarate reductase inhibitors produced by Penicillium sp. FKI-3389 (2011)
  • Author(s): Mori M, Morimoto H, Kim YP, Ui H, Nonaka K, Masuma R, Sakamoto K, Kita K, Tomoda H, Shiomi K, Omura S
  • Journal Title: Tetrahedron Volume: 67 Pages: 6582-6586
• [Journal Article] Uklactones A and B, new NADH-fumarate reductase inhibitors produced by Penicillium sp.FKI-3389 (2011)
  • Author(s): M.Mori, H.Morimoto, Y.-P.Kim, H.Ui, K.Nonaka, R.Masuma, K.Sakamoto, K.Kita, H.Tomoda, K.Shiomi, S.Omura
  • Journal Title: Tetrahedron Volume: 67 Issue: 35 Pages: 6582-6586
  • DOI: 10.1016/j.tet.2011.05.090
  • Peer Reviewed
• [Journal Article] Age-related changes in the activities of respiratory chain complexes and mitochondrial morphology in Drosophila. (2011)
  • Author(s): Oda, Y., Yui, R., Sakamoto, K., Kita, K., Matsuura, E.T.
  • Journal Title: Mitochondrion Volume: 12 Issue: 2 Pages: 345-351
  • DOI: 10.1016/j.mito.2012.01.004
  • Peer Reviewed
• [Journal Article] Trypanosome alternative oxidase, a potential therapeutic target for sleeping sickness, is conserved among Trypanosoma brucei subspecies (2010)
  • Author(s): Nakamura, K., Fujioka, S., Fukumoto, S., Inoue, N., Sakamoto, K., Hirata, H., Kido, Y., Yabu, Y., Suzuki, T., Watanabe, Y., Saimoto, H., Akiyama, H. and Kita, K.
  • Journal Title: Parasitology International Volume: 59 (4) Issue: 4 Pages: 560-564
  • DOI: 10.1016/j.parint.2010.07.006
  • NAID: 10029826326
  • Peer Reviewed
• [Presentation] Rhodobacter capsulatusとRhodospirillum rubrum間でのキメラ型コハク酸‐ユビキノン還元酵素の作製 (2013)
  • Author(s): 福士実咲・柴谷恵太・Hendri Aldrat・北潔・Fevzi Daldal・坂元君年
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜・横浜
• [Presentation] Rhodobacter capsulatusとRhodospirillum rubrum間でのキメラ型コハク酸‐ユビキノン還元酵素の作製 (2013)
  • Author(s): 福士 実咲、柴谷 恵太、Hendri Aldrat、北 潔、Fevzi Daldal、坂元 君年
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] 紅色光合成細菌Rhodobacter capsulatusを用いたコハク酸脱水素酵素複合体の異種生物間発現 (2012)
  • Author(s): 坂元君年・Hendri Aldrat・北潔
  • Organizer: 日本農芸化学会2012年度大会
  • Place of Presentation: 京都女子大学
• [Presentation] コハク酸脱水素酵素複合体の異種生物間発現系の開発 (2011)
  • Author(s): 坂元君年・Hendri Aldrat・北潔
  • Organizer: 日本農芸化学会東北支部第146回大会
  • Place of Presentation: 山形大学
• [Presentation] コハク酸脱水素酵素複合体の異種生物間発現系の開発
  • Author(s): 坂元 君年、Hendri Aldrat、北 潔
  • Organizer: 日本農芸化学会東北支部
  • Place of Presentation: 山形大学農学部
• [Presentation] 紅色光合成細菌Rhodobacter capsulatusを用いたコハク酸脱水素酵素複合体の異種生物間発現
  • Author(s): 坂元 君年、Aldrat Hendri、北 潔
  • Organizer: 日本農芸化学会
  • Place of Presentation: 京都女子大学