Analysis of activation mechanism of new members of NADPH oxidase family

• Project/Area Number: 19590278
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kobe University
• Principal Investigator: UEYAMA Takehiko Kobe University, 自然科学系先端融合研究環バイオシグナル研究センター, 講師 (80346254)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2007: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 活性酸素 / NADPH oxidase / 感染症

Research Abstract

近年、上皮細胞における活性酸素の産生が報告され、続いて組織特異的に存在する新規NADPH oxidase(Nox: 7種類) が確認され、その生理機能やその機能異常によって引き起こされる疾患などに注目が集っている。本研究では、新規Noxの活性化機構の詳細を、遺伝子操作マウスや操作マウス由来の細胞などを用いて解明することを試みた。
その結果、
1. 生細胞を用いて、共焦点レーザー顕微鏡下で、可視化により蛋白質相互作用を検知できる新規蛍光蛋白質システム(complementation-based method using a monomeric coral fluorescent protein: mKG system)を開発し、その有用性をNox複合体の相互作用を用いて証明し、報告した。
2. 遺伝子操作マウス由来の初代培養細胞とレンチウイルスを用いた Nox の再構築系の確立に成功した。

Research Products

• [Journal Article] Sequential binding of cytosolic phox proteins to phagosomes through regulated adaptor proteins: evaluation using the novel monomeric Kusabira-Green system and live imaging of phagocytosis (2008)
  • Author(s): Ueyama T, Kusakabe T, Karasawa S, Kawasaki T, Shimizu A, Son J, Leto TL, Miyawaki A, Saito N
  • Journal Title: J Immunol. 181 Pages: 629-640
• [Journal Article] Enzymological analysis of mutant protein kinase C gamma causing spinocerebellar ataxia type 14 and dysfunction in Ca2+ homeostasis (2008)
  • Author(s): Adachi N, Kobayashi T, Takahashi H, Kawasaki T, Shirai Y, Ueyama T, Matsuda T, Seki T, Sakai N, Saito N.
  • Journal Title: J Biol Chem, 283 Pages: 19854-19863
• [Journal Article] Regulation of clathrin dependent endocytosis by diacylglycerol kinase delta-importance of kinase activity and binding to AP2alpha (2008)
  • Author(s): Kawasaki T, Kobayashi T, Ueyama T, Shirai Y, Saito N
  • Journal Title: Biochem J 409 Pages: 471-479
• [Journal Article] Ca2+ oscillation induced by P2Y2 receptor activation and its regulation by a neuron-specific subtype of PKC (・PKC) (2008)
  • Author(s): Ashida N, Ueyama T, Rikitake K, Shirai Y, Eto M, Kondoh T, Kohmura E, Saito N
  • Journal Title: Neurosci Lett 446 Pages: 123-128
• [Journal Article] 新規アダプター蛋白質(Noxo1, Noxa1)の構造と機能 (2008)
  • Author(s): 上山健彦, 齋藤尚亮
  • Journal Title: G.I. Research(先端医学社) 16 Pages: 208-213
• [Journal Article] Sequential binding of cytosolic phox proteins to phagosomes through regulated adaptor proteins : evaluation using the novel monomeric Kusabira-Green system and live imaging of phagocytosis. (2008)
  • Author(s): Uevama T, Kusakabe T, Karasawa S, Kawasaki T, Shimizu A, Son J, LetoTL, Miyawaki A, Saito N
  • Journal Title: j of Immunology 181 Pages: 629-640
  • Peer Reviewed
• [Journal Article] Enzymological analysis of mutant protein kinase C gamma causing spinocerebeliar ataxia type 14 and dysfunction in Ca^<2+> homeostasis. (2008)
  • Author(s): Adachi N, Kobayashi T, Takahashi H, Kawasaki T, Shirai Y, Ueyama T, Matsuda T, Seki T, Sakai N, Saito N
  • Journal Title: j of Biological Chemistry 283 Pages: 19854-19863
  • Peer Reviewed
• [Journal Article] Regulation of clathrin dependent endocytosis by diacylglycerol kinase delta-importance of kinase activity and binding to AP2alpha. (2008)
  • Author(s): Kawasaki, Kobayashi, Ueyama, Shirai, Saito
  • Journal Title: Biochemical Journal 409 Pages: 471-479
  • Peer Reviewed
• [Presentation] Nox2 活性化時におけるphox蛋白複合体の食胞膜へのターゲティング機構 (2008)
  • Author(s): 上山健彦, 他8名
  • Organizer: 第81回日本薬理学会年会
  • Place of Presentation: パシヒィコ横浜
  • Year and Date: 2008-03-19
• [Presentation] Nox2活性化時におけるphox蛋白複合体の食胞膜へのターゲティング機構 (2008)
  • Author(s): 上山健彦, 他8名
  • Organizer: 第81回日本薬理学会年会
  • Place of Presentation: バシフィコ横浜
  • Year and Date: 2008-03-19
• [Presentation] Sequential binding of cytosolic phox complex to phagosomes through regulated adaptor proteins (2008)
  • Author(s): Ueyama T, Kusakabe T, Karasawa S, Kawasaki T, Shimizu A, Son J, Leto TL, Miyawaki A, Saito N
  • Organizer: Gordon Research Conferences (NADPH oxidase)
  • Place of Presentation: Colby-Sawyer College, NH, USA
• [Presentation] Sequential binding of cytosolic phox complex to phagosomes through regulateda daptor proteins : evaluation using the novel monomeric Kusabira-Green (mKG) system and live imaging of phagocytosis. (2008)
  • Author(s): Ueyama T, Kusakabe T, Karasawa S, Kawasaki T, Shimizu A, SonJ, Leto TL, Miyawaki A, Saito N
  • Organizer: Gordon Research Conferences (NADPH oxidase)
  • Place of Presentation: Colby-Sawyer College, Laconia, NH, USA
• [Presentation] A regulated adaptor function of p40phox: Intramolecular interaction (PX-PB1 domain) within p40phox (2007)
  • Author(s): Ueyama T, Tatsuno T, Kawasaki T, Leto TL, Saito N
  • Organizer: Gordon Research Conferences (Phagocytes)
  • Place of Presentation: Bryant University, Smithfield, RI, USA