Dynamic analysis of photoprotein by using photoaffinity labeling and mass spectrometric analysis

• Project/Area Number: 19780087
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Bioproduction chemistry/Bioorganic chemistry
• Research Institution: Nagoya University
• Principal Investigator: KUSE Masaki Nagoya University, 物質科学国際研究センター, 助教 (40335013)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,270,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥270,000); Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2007: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: 生物発光 / 発光タンパク / 光親和性標識 / 質量分析 / アジド基 / 酸化修飾 / 発光素子

Research Abstract

発光タンパクが効率よく発光している分子機構を明らかにすることを目的として、イクオリンを光親和性標識した。既に合成したアジド化デヒドロセレンテラジンを用いてイクオリンを再構成し、発光前後において光照射した。トリプシン消化した後、質量分析した結果、システイン残基が特異的に酸化修飾されていることが判明した。この現象はこれまでに報告例が無く、発光前後におけるタンパクの動的変化解析に利用できる手法を確立できた。

Research Products

• [Journal Article] Selective oxidation by the hydroperoxide in the photoprotein, aequorin. (2009)
  • Author(s): Doi, I.; Kuse, M.; Nishikawa, T.; Isobe, M.
  • Journal Title: Bioorg. Med. Chem. 17 Pages: 3399-3404
  • Peer Reviewed
• [Journal Article] Differentiation of coelenteramide sulfate from phosphate by hydrogen/ deuterium exchange coupled with ion trap mass spectrometry. (2009)
  • Author(s): Doi, I.; Kuse, M.; Nakashima, Y.; Tani, N.; Isobe, M.
  • Journal Title: J. Mass Spectrom. Soc. Jpn. 57 Pages: 89-95
  • NAID: 10024837397
  • Peer Reviewed
• [Journal Article] Substituent effect of imino-O-arenesulfonates, a coupling partner in Suzuki-Miyaura Reaction for substitution of the Pyrazine Ring: A Study for the Synthesis of Coelenterazine Analogs. (2009)
  • Author(s): Makarasen, A.; Kuse, M.; Nishikawa, T.; Isobe, M.
  • Journal Title: Bull. Chem. Soc. Jpn. 82(in press)
  • NAID: 10025126416
  • Peer Reviewed
• [Journal Article] ^1H NMR Aided Elucidation of Products Derived from Photodegradation of Ethyl 3-azido-4,6-difluorobenzoate in 2,2,2-Trifluoroethanol. (2009)
  • Author(s): Sydnes, M.O.; Kuse, M.; Doi, I.; Isobe, M.
  • Journal Title: Tetrahedron 65 Pages: 3863-3870
  • Peer Reviewed
• [Journal Article] Chemistry for biology of symplectin bioluminescence with fluoro-dehydrocoelenterazine. (2009)
  • Author(s): Nakashima, Y.; Kongjinda, V.; Tani, N.; Kuse, M.; Isobe, M.
  • Journal Title: Chemiluminescence and Bioluminescence Pages: 51-54
  • Peer Reviewed
• [Journal Article] Selective oxidation by the hydroperoxide in the photoprotein, aequorin (2009)
  • Author(s): Doi, I. ; Kuse, M. ; Nishikawa, T. ; Isobe, M.
  • Journal Title: Bioorg., Med. Chem. 17(印刷中)
  • Peer Reviewed
• [Journal Article] Differentiation of coelenteramide sulfate from phosphate by hydrogen/deuterium exchange coupled with ion trap mass spectrometry (2009)
  • Author(s): Doi, I. ; Kuse, M. ; Nakashima, Y. ; Tani, N. ;Isobe, M.
  • Journal Title: J. Mass Spectrom. Soc. Jpn. 57 Pages: 89-95
  • NAID: 10024837397
  • Peer Reviewed
• [Journal Article] Substituent effect of imino-O-arenesulfonates, a coupling partner in Suzuki-Miyaura Reaction for substitution of the Pyrazine Ring : A Study for the Synthesis of Coelenterazine Analogs (2009)
  • Author(s): Makarasen, A. ; Kuse, M. ; Nishikawa, T. ; Isobe, M.
  • Journal Title: Bull. Chem. Soc. Jpn. 82(印刷中)
  • NAID: 10025126416
  • Peer Reviewed
• [Journal Article] Pholasin luminescence is enhanced by addition of dehydrocoelenterazine. (2008)
  • Author(s): Kuse, M.; Tanaka, E.; Nishikawa, T.
  • Journal Title: Bioorg. Med. Chem. Lett. 18 Pages: 5657-5659
  • Peer Reviewed
• [Journal Article] Cysteine-390 is the binding site of luminous substance with symplectin, a photoprotein from Okinawan squid, Symplectoteuthis oualaniensis. (2008)
  • Author(s): Isobe, M.; Kuse, M.; Tani, N.; Fujii, T.; Matsuda, T.
  • Journal Title: Proc. Japan Acad. Ser. B. 84 Pages: 386-392
  • NAID: 130000093921
  • Peer Reviewed
• [Journal Article] Determination of Solvent-Trapped Products Derived from Photolysis of Aryl Azides in 2,2,2-Trifluoroethanol. (2008)
  • Author(s): Sydnes, M.O.; Doi, I.; Ohishi, A.; Kuse, M.; Isobe, M.
  • Journal Title: Chemistry -An Asian J. 3 Pages: 102-112
  • Peer Reviewed
• [Journal Article] Protein Phosphatase Inhibitory Activity of Tautomycin Photoaffinity Probes Evaluated at Femto-molar Level. (2008)
  • Author(s): Sydnes, M.O.; Kuse, M.; Kurono, M.; Shimomura, A.; Ohinata, H.; Takai, A.; Isobe, M.
  • Journal Title: Bioorg. Med. Chem. 16 Pages: 1747-1755
  • Peer Reviewed
• [Journal Article] Reductive monoalkylation of nitro aryls in one-pot. (2008)
  • Author(s): Sydnes, M.O.; Kuse, M.; Isobe, M.
  • Journal Title: Tetrahedron 64 Pages: 6406-6414
  • Peer Reviewed
• [Journal Article] Determination of Solvent-Trapped Products Derived from Photolysis of Aiyl Azides in 2,2,2-Trifluoroethanol. (2008)
  • Author(s): Sydnes, M. O.; Doi, I.; Ohishi, A.; Kuse, M.: Isobe, M.
  • Journal Title: Chemistry-An Asian Journal 3 Pages: 102-112
  • Peer Reviewed
• [Journal Article] Protein Phosphatase Inhibitory Activity of Tautomycin Photoaffinity Probes Evaluated at Femto-molar Level. (2008)
  • Author(s): Sydnes, M. O.; Kuse, M.; Kurono, M.; Shimomura, A.; Ohinata, H.; Takai, A.; Isobe, M.
  • Journal Title: Bioorganic Medicinal Chemistry 16 Pages: 1747-1755
  • Peer Reviewed
• [Presentation] ヒカリカモメガイ発光タンパク(pholasin)の有機分子に関する研究 (2009)
  • Author(s): 田中瑛子、土井一生、久世雅樹、西川俊夫
  • Organizer: 日本農芸化学会2009 年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] ジフルオロデヒドロセレンテラジンを用いたsymplectin発光機構の動的解析 (2009)
  • Author(s): 中島陽介、久世雅樹、谷直紀、西川俊夫、磯部稔
  • Organizer: 日本農芸化学会2009年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] H/D交換質量分析を用いたニトレンの溶媒挿入付加物の解析 (2009)
  • Author(s): 土井一生、Magne Olav Sydnes、久世雅樹、西川俊夫、磯部稔
  • Organizer: 日本農芸化学会2009 年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] Study of di-fluoro-dehydrocoelenterazine analogs in bioluminescence of symplectin (2009)
  • Author(s): Kongjinda, V.; Nakashima, Y.; Tani, N.; Kuse, M.; Nishikawa, T.; Isobe, M.
  • Organizer: 日本農芸化学会2009 年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] ヒカリカモメガイ発光タンパク(pholasin)の有機分子に関する研究 (2009)
  • Author(s): 田中瑛子、土井一生、久世雅樹、西川俊夫
  • Organizer: 日本農芸化学会2009年度大会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] Marine bioluminescence with coelenterate compounds. (2009)
  • Author(s): Masaki Kuse
  • Organizer: The 8^<th> Joint Seminar JSPS-NRCT Core University Program.
  • Place of Presentation: Bangkok
• [Presentation] 化学合成発光素子によるトビイカ発光タンパクsymplectinの生物有機化学的研究 (2008)
  • Author(s): 中島陽介、久世雅樹、西川俊夫、磯部稔
  • Organizer: 日本農芸化学会第153回中部支部例会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-11-01
• [Presentation] 二枚貝発光タンパク(pholasin)の有機基質に関する研究 (2008)
  • Author(s): 田中瑛子、久世雅樹、西川俊夫
  • Organizer: 日本農芸化学会 第153回中部支部例会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-11-01
• [Presentation] オワンクラゲ生物発光機構に関する生物有機化学的研究 (2008)
  • Author(s): 土井一生、久世雅樹、西川俊夫、磯部稔
  • Organizer: 第50回天然物有機化合物討論会
  • Place of Presentation: 福岡
  • Year and Date: 2008-10-02
• [Presentation] Molecular Mechanism of jellyfish bioluminescence by means of photolysis of aequorin. (2008)
  • Author(s): Doi, I.; Kuse, M.; Nishikawa, T.; Isobe, M.
  • Organizer: 15^<TH> INTERNATIONAL SYMPOSIUM ON BIOLUMINESCENCE & CHEMILUMINESCENCE
  • Place of Presentation: Shanghai (China)
  • Year and Date: 2008-05-15
• [Presentation] Chemistry for biology of symplectin bioluminescence with fluoro-dehydrocoelenterazine. (2008)
  • Author(s): Nakashima, Y.; Kongjinda, V.; Tani, N.; Kuse, M.; Isobe, M.
  • Organizer: 15^<TH> INTERNATIONAL SYMPOSIUM ON BIOLUMINESCENCE & CHEMILUMINESCENCE
  • Place of Presentation: Shanghai (China)
  • Year and Date: 2008-05-15
• [Presentation] 光親和性標識によるデプシペプチドへの官能基導入 (2008)
  • Author(s): 大石亜矢子、Makarasen, Arthit、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会2008年度大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-03-27
• [Presentation] オワンクラゲ発光機構に関する生物有機化学的研究 (2008)
  • Author(s): 土井一生、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会2008 年度大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-03-27
• [Presentation] ジフルオロデヒドロセレンテラジンを用いたトビイカ短寿命発光中間体に関する立体過程 (2008)
  • Author(s): 中島陽介、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会2008 年度大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-03-27
• [Presentation] トビイカ発光タンパク質シンプレクチンの発光素子結合部位の探索 (2008)
  • Author(s): 谷直紀、長瀬さやか、中島陽介、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会2008 年度大会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-03-27
• [Presentation] FLUORO-DEHYDROCOELENTERAZINE: CHEMICAL BIOLOGY OF SYMPLECTIN. (2007)
  • Author(s): Kuse, M.; Nakashima, Y.; Tani, N.; Doi, I.; Isobe, M.
  • Organizer: Princess Congress VI
  • Place of Presentation: タイ王国・バンコク
  • Year and Date: 2007-11-26
• [Presentation] ジフルオロデヒドロセレンテラジンを用いたトビイカ短寿命発光中間体に関する立体過程 (2007)
  • Author(s): 中島陽介、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会関西支部・中部支部合同大会
  • Place of Presentation: 名古屋
  • Year and Date: 2007-09-22
• [Presentation] K^+選択的イオノフォア機能解析のためのフォトプローブ分子設計 (2007)
  • Author(s): 大石亜矢子、Arthit Makarasen、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会関西支部・中部支部合同大会
  • Place of Presentation: 名古屋
  • Year and Date: 2007-09-22
• [Presentation] Fluoro-Dehydrocoelenterazines: Chemical Biology of Symplectin Bioluminescence. (2007)
  • Author(s): Kuse, M.; Nakashima, Y.; Tani, N.; Doi, I.; Isobe, M.
  • Organizer: 2nd International Conference of Cutting-Edge Organic Chemistry in Asia.
  • Place of Presentation: 韓国・釜山
  • Year and Date: 2007-09-04
• [Presentation] フッ素2置換デヒドロセレンテラジンの物性と生物発光 (2007)
  • Author(s): 中島陽介、久世雅樹、磯部稔
  • Organizer: 日本農芸化学会2007年度大会
  • Place of Presentation: 東京
  • Year and Date: 2007-03-26
• [Presentation] FLUORODEHYDROCOELENTERAZINE : CHEMICAL BIOLOGY OF SYMPLECTIN. (2007)
  • Author(s): Kuse, M.; Nakashima, Y.; Tani, N.; Doi, I.; Isobe, M.
  • Organizer: Princess Congress VI
  • Place of Presentation: Bangkok, Thailand
• [Presentation] Photoaffinity labeling of Aequorin with an Azide-coelenterazine. (2007)
  • Author(s): Doi, I.; Kuse, M.; Isobe, M.
  • Organizer: Princess Congress VI
  • Place of Presentation: Bangkok, Thailand
• [Presentation] Fluoro-Dehydrocoelenterazines: Chemical Biology of Symplectin Bioluminescence. (2007)
  • Author(s): Kuse,M.; Nakashima, Y.; Tani, N.; Doi, I.; Isobe, M.
  • Organizer: 2^<nd> International Conference of Cutting-Edge Organic Chemistry in Asia.
  • Place of Presentation: Pusan, Korea
• [Remarks]
  • URL: http://www.cic.nagoya-u.ac.jp/