Elucidation of the functional regulation of myristoylated proteins in intracellular signaling network

• Project/Area Number: 19570139
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: Kyoto Gakuen University
• Principal Investigator: MATSUBARA Mamoru Kyoto Gakuen University, バイオ環境学部, 准教授 (90288481)
• 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: ミリスチル化 / 翻訳後修飾 / 細胞内シグナル伝達 / リン酸化 / プロテオーム解析プロテインキナーゼ / プロテオーム / プロテインキナーゼ / 質量分析 / シグナル伝達 / 蛋白質相互作用

Research Abstract

細胞内シグナルネットワークにおけるミリスチル化蛋白質の機能調節機構を理解するために、NAP-22についてミリスチル化依存的に結合する蛋白質の同定を行った。その結果、従来知られているカルモジュリン以外にPKCαやERK1, 2などのプロテインキナーゼが同定された。ERK1とNAP22が直接結合すると同時に、直接リン酸化することも分かった。更に、リン酸化部位同定法を新たに開発し、これらERK1でリン酸化される部位を正確に同定した。

Research Products

• [Journal Article] Biochemical characterization of Rab proteins from Bombyx mori (2009)
  • Author(s): Uno, T., Morikawa T., Nakamura, M., Matsubara, M., Yamagata, H., Kanamaru, K. and Takagi, M.
  • Journal Title: Arch. Insect Biochem. Physiol 70 Pages: 77-89
  • Peer Reviewed
• [Journal Article] Two-dimensional phosphate-affinity gel electrophoresis for the analysis of phoshoprotein isotypes (2009)
  • Author(s): Kinoshita E., Kinoshita-Kikuta E., Matsubara, M., Aoki Y., Ohie, S., Mouri, Y. and Koike T.
  • Journal Title: Electrophoresis 30 Pages: 550-559
  • Peer Reviewed
• [Journal Article] Two-dimensional phosphate-affinity gelelectrophoresis for the analysis of phoshoprotein isotypes. (2009)
  • Author(s): Kinoshita, E.
  • Journal Title: Electrophoresis 30 Pages: 550-559
  • Peer Reviewed
• [Journal Article] Biochemical characterization of Rab proteias from Bombyxmorl. (2009)
  • Author(s): Uno, T.
  • Journal Title: Arch. Insect Biochem. Physiol. 70 Pages: 77-89
  • Peer Reviewed
• [Journal Article] Neural complex-specific expression of xylosyl N-glycan in Ciona intestinalis (2008)
  • Author(s): Yagi, H., Nakagawa, M., Takahashi, N., Kondo, S., Matsubara, M. and Kato, K.
  • Journal Title: Glycobiology 18 Pages: 145-151
  • Peer Reviewed
• [Journal Article] Separation of phosphoproteins isotypes having the same number phosphate groups using phoshate-affinity SDS-PAGE (2008)
  • Author(s): Kinoshita, E., Kinoshita-Kikuta, Matsubara, M, Yamada, S., Nakamura, H., Shiro, Y. Aoki, Y., Okita, K. and Koike, T.
  • Journal Title: Proteomics 15 Pages: 2994-3003
  • Peer Reviewed
• [Journal Article] Crystal structure of human mono-phosphorylated ERK1 at Tyr204 (2008)
  • Author(s): Kinoshita, T., Yoshida, I., Nakase, S., Okita, K., Gouda, M., Matsubara, M., Yokota, K., Ishiguro, H. and Tada, T
  • Journal Title: Biochem. Biophys. Res. Communu 377 Pages: 1123-1127
  • Peer Reviewed
• [Journal Article] The neuronal connexin36 interacts with and is phosphorylated by CaMKII in a way similar to CaMKII interaction with glutamate receptors (2008)
  • Author(s): Alev, C., Urschel, S., Sonntag, S., Zoidl, G., Fort, A., Hoher, T., Matsubara, M., Willecke, K., Spray, D. and Dermietzel, R.
  • Journal Title: Proc. Nat. Acad. Sci. U.S.A 105 Pages: 20964-20969
  • Peer Reviewed
• [Journal Article] Separat ion of phosphoproteins isotypes having the same number phosphate groups using phosphate-affinity SDS-PAGE. (2008)
  • Author(s): Kinoshi ta, E.
  • Journal Title: Proteomics 15 Pages: 2994-3003
  • Peer Reviewed
• [Journal Article] Crystal strueture of human mono-phosphorylated ERK1 at Tyr204. (2008)
  • Author(s): Kinoshita, T.
  • Journal Title: Biochem. Biophys. Res. Communu. 377 Pages: 1123-1127
  • Peer Reviewed
• [Journal Article] The neuronal connexin36 interacts with and is phosphorylated by CaMKll in a way similar to CaMKII interaction with glutamate receptors. (2008)
  • Author(s): Alev, C.
  • Journal Title: Proc. Nat. Acad. Sci. U.S.A. 105 Pages: 20964-20969
  • Peer Reviewed
• [Journal Article] Neural complex-specific expression of xylosyl N-glycan in Ciona intestinalis. (2008)
  • Author(s): Yagi, H.
  • Journal Title: Glycobiology 18 Pages: 145-151
  • Peer Reviewed
• [Journal Article] プロテオミクスによる環境応答シグナル伝達機構の解明 (2008)
  • Author(s): 松原 守
  • Journal Title: 京都学園大学 総合研究所所報 9 Pages: 59-63
• [Journal Article] Determination of phosphorylated amino acid residues of Rab8 from Bombyx mori (2007)
  • Author(s): Uno, T., Nakada, T., Okamaoto, S., Nakamura, M., Matsubara, M., Imaishi, H., Yamagata, H., Kanamaru, K. and Takagi, M.
  • Journal Title: Arch. Insect Biochem. Physiol 66 Pages: 89-97
  • Peer Reviewed
• [Journal Article] Dynamic site-specific phosphorylation analysis by LC/MS/MS (2007)
  • Author(s): Matsubara, M., Hunter, C. and Tsuchiya, F.
  • Journal Title: Chemistry Today 25 Pages: 92-94
  • Peer Reviewed
• [Journal Article] プロテオミクスによる環境応答シグナル伝達機構の解明 (2007)
  • Author(s): 松原守
  • Journal Title: 京都学園大学総合研究所所報 9 Pages: 59-63
• [Journal Article] Dynamic site-specific phosphorylation analysis by LC/MS/MS. (2007)
  • Author(s): Matsubara, M.
  • Journal Title: Chemistry Today 25 Pages: 92-94
  • Peer Reviewed
• [Journal Article] Determination of phosphorylatin amino acid residues of Rab8 from Bombyx mori. (2007)
  • Author(s): Uno, T.
  • Journal Title: Arch. Insect Biochem. Physiol. 66 Pages: 89-97
  • Peer Reviewed
• [Presentation] リボソームタンパク質S19のリン酸化とS19結合タンパク質との相互作用 (2008)
  • Author(s): 前田紀子、徳誠吉、内藤康仁、松原守、山本秀幸
  • Organizer: BMB2008
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-12
• [Presentation] Separation of phosphoprotein isotypes with the same phosphate stoichiometry using phosphate-affinity SDS-PAGE (2008)
  • Author(s): Eiji Kinoshita, Emiko Kinoshita-Kikuta, Mamoru Matsubara, Seiji Yamada, Hiro Nakamura, Yoshitsugu, Shiro, Yuri Aoki, Kouki, Tohru Koike
  • Organizer: BMB2008
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-12
• [Presentation] Separation of phosphoprotein isotypes with the same phosphate stoichiometry using phosphate-affinity SDS-PAGE. (2008)
  • Author(s): Eiji Kinoshita
  • Organizer: BMB2008
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2008-12-12
• [Presentation] カタユウレイボヤにおける神経複合体特異的なキシロース含有N型糖鎖の発現 (2008)
  • Author(s): 矢木宏和、中川将司、高橋禮子、金子琢磨、近藤幸子、松原守、加藤晃一
  • Organizer: BMB2008
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] カタユウレイボヤの神経複合体におけるキシロース含有N型糖鎖の特異的発現 (2008)
  • Author(s): 矢木宏和、中川将司、近藤幸子、高橋禮子、松原守、加藤晃一
  • Organizer: 日本薬学会第128年会
  • Place of Presentation: 横浜
  • Year and Date: 2008-03-28
• [Presentation] ヒトERK1/Iodotubercidin複合体の構造学的研究 (2008)
  • Author(s): 吉田育代、木下誉富、合田正貴、横田耕一、松原守、石黒啓司、多田俊治
  • Organizer: 日本薬学会第128年会
  • Place of Presentation: 横浜
  • Year and Date: 2008-03-26
• [Presentation] Phosphate affinity 2-D gel electrophoresis for the analysis of Phosphoproteins (2007)
  • Author(s): Eiji Kinoshita, Emiko Kinoshita-Kikuta, Yuri Aoki, Shiori Ohie, Yuka Mouri, Mamoru Matsubara, and Tohru Koike
  • Organizer: BMB2007
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-13
• [Presentation] 質量分析計を用いたリン酸化ネットワークの解析 (2007)
  • Author(s): 松原守
  • Organizer: 第3回(2007)島津製作所MALDIユーザーズミーティング基調講演
  • Place of Presentation: 大阪
  • Year and Date: 2007-07-27
• [Presentation] 質量分析計を用いたリン酸化ネットワークの解析(招待講演) (2007)
  • Author(s): 松原 守
  • Organizer: 第3回島津製作所 MALDIユーザーズミーティング
  • Place of Presentation: 大阪
  • Year and Date: 2007-07-27
• [Presentation] リン酸化結合ナノ分子を利用した新しい2次元電気泳動法の開発 (2007)
  • Author(s): 木下英司、木下恵美子、大家志織、松原守、小池透
  • Organizer: 第48回日本生化学会中国・四国支部例会
  • Place of Presentation: 高知
  • Year and Date: 2007-05-20
• [Presentation] リン酸化プロテオーム研究に向けた新しいリン酸アフィ二ティー2次元ゲル電気泳動法の開発 (2007)
  • Author(s): 大家志織、松原守、青木悠里、木下英司、木下恵美子、小池透
  • Organizer: 日本薬学会第127年会
  • Place of Presentation: 富山
  • Year and Date: 2007-03-29
• [Presentation] Monitoring of temporal changes in phosphorylation states of proteins using mass spectrometry and a chemical labeling strategy (2007)
  • Author(s): Tsuchiya, F., Hunter, C. and Matsubara, M.
  • Organizer: ABRF2007-Creating the Biological Roadmap-
  • Place of Presentation: Tampa, Florida
• [Remarks]
  • URL: http://www.kyotogakuen.ac.jp/~molbio/
• [Remarks]
  • URL: http://www.kyotogakuen.ac.jp/~molbio/index.html
• [Remarks]
  • URL: http://homepage3.nifty.com/m_matsubara/index.html
• [Remarks]
  • URL: http://www.kyotogakuen.ac.jp/~molbio/