X-ray and neutron crystallographic structure of copper amine oxidase

• Project/Area Number: 26440037
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Osaka Medical College
• Principal Investigator: Murakawa Takeshi 大阪医科大学, 医学部, 助教 (90445990)
• Co-Investigator(Kenkyū-buntansha): 林 秀行 大阪医科大学, 医学部, 教授 (00183913)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 中性子結晶構造解析 / X線結晶構造解析 / トパキノン / 酵素 / 中性子結晶構造回析 / X線結晶構造回析 / 中性子 / 補酵素 / Ｘ線

Outline of Final Research Achievements

Copper amine oxidases (CAOs) catalyze the oxidation of various aliphatic amines to the corresponding aldehydes, ammonia and hydrogen peroxide. We focused on the structural insights into the catalytic mechanism of a CAO from Arthrobacter globiformis (AGAO) using an ultra-high resolution X-ray and neutron crystallography. We determined the structure of oxidized-form (substrate-free form) of AGAO at 1.72/1.14 angstrom　by neutron/X-ray crystallography. Based on the precise three-dimensional structure information obtained, we got new knowledge on reaction mechanism accompanied by structural change and fluctuation of AGAO. Further, we also obtained the neutron and X-ray diffraction data of semiquinone intermediate. We are currently refining the structure.

Research Products

• [Journal Article] Probing the Catalytic Mechanism of Copper Amine Oxidase from Arthrobacter globiformis with Halide Ions. (2015)
  • Author(s): Murakawa, T., Hamaguchi, A., Nakanishi, S., Kataoka, M., Nakai, T., Kawano, Y., Yamaguchi, H., Hayashi, H., Tanizawa, K., and Okajima, T.
  • Journal Title: J. Biol. Chem. Volume: 290 Issue: 38 Pages: 23094-23109
  • DOI: 10.1074/jbc.m115.662726
  • Peer Reviewed
• [Presentation] 銅含有アミン酸化酵素における触媒反応中のプロトン化状態についての理論的解明 (2017)
  • Author(s): 庄司光男，村川武志，重田育照，岡島俊英
  • Organizer: 第１７回日本蛋白質科学会年会
• [Presentation] X-ray crystallographic structure of semiquinone radical intermediate formed in bacterial copper amine oxidase. (2016)
  • Author(s): Toshihide Okajima, Tadashi Nakai1 , Katsuyuki Tanizawa , Takeshi Murakawa , and Hideyuki Hayashi.
  • Organizer: ICC05-AEM2016
  • Place of Presentation: Unazuki, Japan.
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] 2.Analysis of the Catalytic Mechanism of Copper Amine Oxidase from Arthrobacter Globiformis. (2016)
  • Author(s): Hiroshi Yamaguchi, Takeshi Murakawa, Misumi Kataoka, Yoshiaki Kawano, Hideyuki Hayashi, Katsuyuki Tanizawa, and Toshihide Okajima.
  • Organizer: ISDSB2016
  • Place of Presentation: Tennessee, USA.
  • Year and Date: 2016-08-07
  • Int'l Joint Research
• [Presentation] 銅含有アミン酸化酵素の大型結晶調製と予備的中性子回折実験 (2015)
  • Author(s): 村川武志
  • Organizer: 茨城県平成２７年度 第３回(第11 回) iBIX 研究会
  • Place of Presentation: 茨城県東海村
  • Year and Date: 2015-10-28
  • Invited
• [Presentation] 銅アミン酸化酵素のTPQ補酵素とその前駆体Tyr残基のコンフォメーション制御の分子機構 (2014)
  • Author(s): 村川武志，中西将太，元山暁仁，林秀行，谷澤克行，岡島俊英
  • Organizer: 日本生化学会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Conformational flexibility of the topa quinone cofactor in copper amine oxidase revealed by site-specific mutagenesis of the neighboring conserved asparagine residue (2014)
  • Author(s): Takeshi Murakawa, Hideyuki Hayashi, Katsuyuki Tanizawa, Toshihide Okajima
  • Organizer: ICC-04
  • Place of Presentation: イタリア，パルマ
  • Year and Date: 2014-08-24 – 2014-08-30
• [Presentation] 銅アミン酸化酵素のTPQ生成反応と触媒反応における保存性Asn残基の役割 (2014)
  • Author(s): 村川武志，中西将太，元山暁仁，林秀行，谷澤克行，岡島俊英
  • Organizer: 日本蛋白質科学会
  • Place of Presentation: 横浜
  • Year and Date: 2014-06-25 – 2014-06-27