Photofunctionalization of apoptotic enzymes for targeting cancer therapy and its applications

• Project/Area Number: 18510184
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Living organism molecular science
• Research Institution: Kyoto University (2008); Osaka University (2006-2007)
• Principal Investigator: ENDO Masayuki Kyoto University, 物質-細胞統合システム拠点, 准教授 (70335389)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,140,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥540,000); Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2007: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: アポトーシス / カスパーゼ-3 / DNA 断片化因子 / 光化学 / 非天然アミノ酸 / 胞外翻訳反応 / 癌治療 / 細胞導入 / DNA断片化因子 / ガスパーゼー3 / 細胞外翻訳反応 / カスパーゼー3

Research Abstract

アポトーシスの誘導による癌細胞の細胞死を誘導することでその除去を目的とし、その間連連酵素であるcaspase-3とDNA断片化因子に光応答性分子を組み込み、光照射によって活性化することで人工的なアポトーシスの誘導を検討した。光分解性の2-nitrophenylglycine4,5-dimethoxy-2-nitrophenylglycine(DMNpg)をDNA断片化因子の阻害タンパク(ICAD)の切断位置に導入し、細胞外転写翻訳反応で光機能化ICADが高収率で発現され、精製した。精製した光機能化DNA断片化因子へ光照射を行うとICADの分解が起こり、光機能化DNA断片化因子複合体への光照射によって、DNase活性の発現を行えることを見出した。さらに細胞内導入のためにHIV由来のTAT配列の導入した光機能化DNA断片化因子を合成した。これによって細胞導入と光照射によるアポトーシスの誘導を検討できる系を構築した。

Research Products

• [Journal Article] DNA Supramolecular Structures and Nanostructures for the Creation of Functional Nanomaterials (2009)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Soft Nanomaterials (in press)
• [Journal Article] DNA Supramolecular Structures and Nanostructures for the Creation of Functional Nanomaterials (2009)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Soft Nanomaterials
  • Peer Reviewed
• [Journal Article] Diastereochemically Controlled Porphyrin Dimer Formation on a DNA Duplex Scaffold (2008)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: J. Org. Chem. 73 Pages: 1106-1112
• [Journal Article] Programmable conformational regulation of porphyrin dimers on geometric scaffold of duplex DNA (2008)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Tetrahedron 64 Pages: 1839-1846
• [Journal Article] Diastereochemically Controlled Porphyrin Dimer Formation on a DNA Duplex Scaffold (2008)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Journal of Organic Chemistry 73 Pages: 1106-1112
  • Peer Reviewed
• [Journal Article] Programmable conformational regulation of porphyrin dimers on geometric scaffold of duplex DNA (2008)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Tetrahedron 64 Pages: 1839-1846
  • Peer Reviewed
• [Journal Article] Porphyrin Light-Harvesting Arrays Constructed in the Recombinant Tobacco Mosaic Virus Scaffold (2007)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Chem. Eur. J. 13 Pages: 8660-8666
• [Journal Article] Monitoring of Three Distinct Structures of Restriction Enzyme Complexes Using Characteristic Fluorescence from Site-selectively Incorporated Solvatochromic Probe (2007)
  • Author(s): K. Nakayama, M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Photochem. Photobiol. Sci. 6 Pages: 836-841
• [Journal Article] Photochemical control of enzymes using photoresponsive compounds on the basis of the interaction mechanism (2007)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Photomed. Photobiol 29 Pages: 14-15
• [Journal Article] Porphyrin Light-Harvesting Arrays Constructed in the Recombinant Tobacco Mosaic Virus Scaffold (2007)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Chemistry, A European Journal 13 Pages: 8660-8666
  • Peer Reviewed
• [Journal Article] Monitoring of Three Distinct Structures of Restriction Enzyme Complexes Using Characteristic Fluorescence from Site-selectively Incorporated Solvatochromic Probe (2007)
  • Author(s): K. Nakayama, M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Photochemical & Photobiological Sciences 6 Pages: 836-841
  • Peer Reviewed
• [Journal Article] Photochemical Properties of Porphyrin-attached Tobacco Mosaic Virus (2007)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: Photomedicine and Photobiology 29 Pages: 19-20
• [Journal Article] Photochemical control of enzymes using photoresponsive compounds on the basis of the interaction mechanism (2007)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Photomedicine and Photobiology 29 Pages: 14-15
• [Journal Article] DNAナノテクノロジー (2007)
  • Author(s): 遠藤政幸
  • Journal Title: 化学フロンティア「ゲノム化学」 18 Pages: 169-177
  • Peer Reviewed
• [Journal Article] Molecular Design of DNA Supramolecular Structures for the Creation of DNA Nanostructures and Functional Nanomaterials (2007)
  • Author(s): M.Endo, T.Majima
  • Journal Title: Soft Nanomaterials (in press)
• [Journal Article] Thermodynamic properties of branched DNA complexes with full-matched and mismatched DNA strands (2006)
  • Author(s): M. Endo, T. Majima
  • Journal Title: Chem. Commun. Pages: 2329-2331
• [Journal Article] Pyrene-Stacked Nanostructures Constructed in the Recombinant Tobacco Mosaic Virus Rod Scaffold (2006)
  • Author(s): M. Endo, H. Wang, M. Fujitsuka, T. Majima
  • Journal Title: Chem. Eur. J. 12 Pages: 3735-3740
• [Journal Article] Detection of the Local Structural Changes in the Dimer Interface of BamHI Initiated by DNA Binding and Dissociation Using a Solvatochromic Fluorophore (2006)
  • Author(s): K. Nakayama, M. Endo, M. Fujitsuka, T. Majima
  • Journal Title: J. Phys. Chem. B 110 Pages: 21311-21318
• [Journal Article] Pyrene-Stacked Nanostructures Constructed in the Recombinant Tobacco Mosaic Virus Rod Scaffold (2006)
  • Author(s): M.Endo, H.Wang, M.Fujitsuka, T.Majima
  • Journal Title: Chemistry,A European Journal 12 Pages: 3735-3740
• [Journal Article] Thermodynamic properties of branched DNA complexes with full-matched and mismatched DNA strands (2006)
  • Author(s): M.Endo, T.Majima
  • Journal Title: Chemical Communications Pages: 2329-2331
• [Journal Article] Detection of the Local Structural Changes in the Dimer Interface of BamHI Initiated by DNA Binding and Dissociation Using a Solvatochromic Fluorophore (2006)
  • Author(s): K.Nakayama, M.Endo, M.Fujitsuka, T.Majima
  • Journal Title: Journal of Physical Chemistry B 110 Pages: 21311-21318
• [Presentation] ポルフィリンを導入したタバコモザイクウィルス超分子の光機能性 (2008)
  • Author(s): 遠藤政幸, 藤塚守, 真嶋哲郎
  • Organizer: 日本化学会第88回年会
  • Place of Presentation: 立教大学・東京
  • Year and Date: 2008-03-29
• [Presentation] 生体超分子を使った機能性分子の自己集積化 (2007)
  • Author(s): 遠藤政幸
  • Organizer: 京都大学大学院理学研究科
  • Year and Date: 2007-12-22
• [Presentation] 活性化機構を基にした酵素の光化学的制御 (2007)
  • Author(s): 遠藤政幸
  • Organizer: 第29回光医学光生物学会シンポジウム「光とバイオテクノロジー」
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2007-07-12
• [Presentation] 光機能化タバコモザイクウィルス超分子 (2007)
  • Author(s): 遠藤政幸, 藤塚守, 真嶋哲朗
  • Organizer: 第29回光医学光生物学会
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2007-07-12
• [Presentation] 活性化機構を基にした酵素の光化学的制御 (2007)
  • Author(s): 遠藤政幸
  • Organizer: 第29回光医学光生物学会
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2007-07-12
• [Presentation] 「光応答性生体超分子の創成」「光と生命」 (2007)
  • Author(s): 遠藤政幸
  • Organizer: 立命館大学草津キャンパス
  • Year and Date: 2007-05-18
• [Presentation] Pyrene-Assmbled Nanostructures Constructed in the Tobacco Mosaic Virus Scaffold (2006)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Organizer: 3rd Korea-Japan Symposium on Frontier Photoscience
  • Place of Presentation: Seoul, Korea,
• [Presentation] DNA Nano-Structures Controlled by Branched DNA Connectors (2006)
  • Author(s): M. Endo, M. Fujitsuka, T. Majima
  • Organizer: 3rd Korea-Japan Symposium on Frontier Photoscience
  • Place of Presentation: Seoul, Korea
• [Presentation] 「精密な分子配列を指向したDNA ナノテクノロジー」シンポジウム (2006)
  • Author(s): 遠藤政幸
  • Organizer: ゲノム化学の最先端-医学・分子生物学への応用と展開-
  • Place of Presentation: 郡山
• [Presentation] ピレンを導入したタバコモザイクウィルス超分子の光化学的性質 (2006)
  • Author(s): 遠藤政幸・藤塚守・真嶋哲朗
  • Organizer: 光化学討論会
  • Place of Presentation: 仙台
• [Presentation] 分岐型DNAによるDNAチューブ構造形成の制御 (2006)
  • Author(s): 遠藤政幸・N. C. Seeman・真嶋哲朗
  • Organizer: 第86 回日本化学会春季年会
  • Place of Presentation: 千葉
• [Presentation] アゾベンゼン誘導体を導入した制限酵素BamHI活性の光異性化による制御 (2006)
  • Author(s): 中山公志・遠藤政幸・真嶋哲朗
  • Organizer: 第86回日本化学会春季年会
  • Place of Presentation: 千葉
• [Presentation] 生体分子高次構造の構築と機能化 (2006)
  • Author(s): 遠藤政幸
  • Organizer: ナノバイオテクノロジーの最先端と新展開
  • Place of Presentation: 兵庫
• [Book] "DNAナノテクノロジー"化学フロンティア「ゲノム化学」化学同人(18巻) (2007)
  • Author(s): 遠藤政幸
• [Book] 化学フロンティア18「ゲノム化学の最前線」 (2007)
  • Author(s): 遠藤政幸
  • Publisher: 化学同人(印刷中)