Analysis of oxygen activation mechanism in the heme-containing dioxygenases

• Project/Area Number: 19770094
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Structural biochemistry
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: SUGIMOTO Hiroshi The Institute of Physical and Chemical Research, 城生体金属科学研究室, 専任研究員 (90344043)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,730,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥330,000); Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2007: ¥2,300,000 (Direct Cost: ¥2,300,000)
• Keywords: ヘムタンパク質 / 共鳴ラマン分光 / X線結晶学 / トリプトファン / 構造生物学

Research Abstract

2つのヘム含有二原子酸素添加酵素(IDO,TDO)の反応メカニズムを明らかにするために共鳴ラマン分光法によりIDOの反応中間体である酸素結合型の電子状態を明らかにした。また、IDO変異体の機能解析・X線結晶構造解析・共鳴ラマン分光解析により明らかにした活性中心のタンパク質残基の役割とDFT法による理論計算結果から、新しい反応スキームを提案した。さらに、TDOの協同的基質結合性とそのメカニズム明らかにした。

Research Products

• [Journal Article] Cooperative binding of L-trp to human tryptophan 2,3-dioxygenase : resonance Raman spectroscopic analysis. (2009)
  • Author(s): Fukumura, E., Sugimoto, H., Misumi, Y., Ogura, T., Shiro, Y.
  • Journal Title: J. Biochem. 145 Pages: 505-515
  • NAID: 10026302198
  • Peer Reviewed
• [Journal Article] Structure-based design of a highly active vitamin D hydroxylase from Streptomyces griseolus CYP105A1. (2008)
  • Author(s): Hayashi, K., Sugimoto, H., Shinkyo, R., Yamada, M., Ikeda, S., Ikushiro, S., Kamakura, M., Shiro, Y., Sakaki, T.
  • Journal Title: Biochemistry 47 Pages: 11964-11972
  • Peer Reviewed
• [Journal Article] Density functional theory study on a missing piece in understanding of heme chemistry : the reaction mechanism for indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase. (2008)
  • Author(s): Chung, L. W., Li, X., Sugimoto, H., Shiro, Y., Morokuma, K.
  • Journal Title: J. Am. Chem. Soc. 130 Pages: 12299-12309
  • Peer Reviewed
• [Journal Article] Synthesis and biological activity of 1-methyl-tryptophan-tirapazamine hybrids as hypoxia-targeting indoleamine 2,3-dioxygenase inhibitors. (2008)
  • Author(s): Nakashima, H., Uto, Y., Nakata, E., Nagasawa, H., Ikkyu, K., Hiraoka, N., Nakashima, K., Sasaki, Y., Sugimoto, H., Shiro, Y., Hashimoto, T., Okamoto, Y., Asakawa, Y., Hori, H.
  • Journal Title: Bioorg. Med. Chem. 16 Pages: 8661-8669
  • Peer Reviewed
• [Journal Article] Crystal structure of CYP105A1 (P450SU-1) in complex with 1alpha,25-dihydroxyvitamin D_3. (2008)
  • Author(s): Sugimoto, H., Shinkyo, R., Hayashi, K., Yoneda, S., Yamada, M., Kamakura, M., Ikushiro, S., Shiro, Y., Sakaki, T.
  • Journal Title: Biochemistry 47 Pages: 4017-4027
  • Peer Reviewed
• [Journal Article] Synthesis and biological activity of 1-methyl-tryptophan-tirapazamine hybrids as hypoxia-targeting indoleamine 2,3-dioxygenase inhibitors (2008)
  • Author(s): Hitomi Nakashima, et al.
  • Journal Title: Bioorganic & Medical Chemistry 16 Pages: 8661-8669
  • Peer Reviewed
• [Journal Article] Density functional theory study on a missing piece in understanding of heme chemistry : the reaction mechanism for indoleamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase (2008)
  • Author(s): Lung Wa Chung, et al.
  • Journal Title: Journal of American Chemical Society 130 Pages: 12299-12309
  • Peer Reviewed
• [Journal Article] Cooperative binding of L-Trp to human tryptophan 2,3-dioxygenase : resonance Raman apectroscopic analysis
  • Author(s): Eiko Fukumura, et al.
  • Journal Title: Journal of Biochemistry (印刷中)
  • Peer Reviewed
• [Journal Article] Crystal structure of CYP105A1 (P450SU-1) in complex with 1□lpha, 25-dihydroxyvitamin D3
  • Author(s): Hiroshi Sugimoto
  • Journal Title: Biochemistry In press
  • Peer Reviewed
• [Presentation] 共鳴ラマン分光法によるトリプトファン2,3-ジオキシゲナーゼの反応機構解析 (2008)
  • Author(s): 福村栄維子、三角裕子、杉本宏、永野真吾、瀧尾擴士、井柳〓、小倉尚志、城宜嗣
  • Organizer: 日本生物物理学会
  • Place of Presentation: 福岡
  • Year and Date: 2008-12-03
• [Presentation] 共鳴ラマン分光法によるトリプトファン2,3-ジオキシゲナーゼの反応機構解析 (2008)
  • Author(s): 福村栄維子, ら
  • Organizer: 第46回日本生物物理学会年会
  • Place of Presentation: 福岡
  • Year and Date: 2008-12-03
• [Presentation] Cytochrome P450SU-1 (CYP105A1)の結晶構造と活性型ビタミンD生成反応のメカニズム (2007)
  • Author(s): 杉本 宏
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-23
• [Presentation] ヒト由来Tryptophan 2,3-dioxygenaseの分光学的解析 (2007)
  • Author(s): 福村栄維子、杉本宏、永野真吾、瀧尾擴士、井柳尭、小倉尚志、城宜嗣
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-21
• [Presentation] 共鳴ラマン分光法によるインドールアミン2,3ジオキシゲナーゼの解析 (2007)
  • Author(s): 四ッ谷景子、杉本宏、大槻崇史、佐藤秀明、吉田匡、小倉尚志、城宜嗣
  • Organizer: 日本生物物理学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-21
• [Presentation] ヒト由来Indoleamine 2,3-dioxygenaseの構造に基づいた変異体解析 (2007)
  • Author(s): 米谷依梨紗、杉本宏、エドワード ヴォッテロ、四ッ谷景子、吉田匡、城宜嗣
  • Organizer: 日本生化学会・日本分子生物学会合同年会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-21
• [Remarks] RIKENリサーチハイライト「Missing piece gets a work over」
  • URL: http://www.rikenresearch.riken.jp/research/627/
• [Remarks]
  • URL: http://www.rikenresearch.riken.jp/research/627/