Functional analysis of the molecular complex in spatiotemporal regulation of protein phosphorylation.

• Project/Area Number: 24592805
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Kyushu Dental College
• Principal Investigator: TAKEUCHI HIROSHI 九州歯科大学, 歯学部, 教授 (70304813)
• Co-Investigator(Renkei-kenkyūsha): GAO Jing 九州大学, 大学院歯学研究院, 助教 (40585882)
• Research Collaborator: ZHANG Zhao ; NAGANO Koki ; WANG Daguang
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: リン酸化 / 脱リン酸化 / 細胞内情報伝達 / 開口分泌 / 細胞内情伝達機構

Outline of Final Research Achievements

We have previously isolated a signaling molecule, PRIP which forms the protein complex with multiple kinases and phosphatases. In this year, we identified that (a)the C2 domains of PRIP and those of other molecules form hetero-dimers, and (b)a PRIP-interacting kinase Akt regulates exocytosis by phosphorylation of its new substrate tomosyn.
The results obtained in this study would promote understanding the mechanism how the phospho-dependent cellular signaling is spatio-temporally regulated by the complex containing multiple enzymes responsible for protein phosphorylation/de-phosphorylation.

Research Products

• [Journal Article] Promotion of insulin-induced glucose uptake in C2C12 myotubes by osteocalcin. (2015)
  • Author(s): Tsuka, S., Aonuma, F., Higashi, S., Ohsumi, T., Nagano, K., Mizokami, A., Kawakubo-Yasukochi, T., Masaki, C., Hosokawa, R. and Hirata, M. and Takeuchi, H.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 459 Issue: 3 Pages: 437-442
  • DOI: 10.1016/j.bbrc.2015.02.123
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Tomosyn is a novel Akt substrate mediating insulin-dependent GLUT4 exocytosis. (2015)
  • Author(s): Nagano, K., Takeuchi, H., Gao, J., Mori, Y., Otani, T., Wang, D. and Hirata, M.
  • Journal Title: The International Journal of Biochemistry & Cell Biology Volume: 62 Pages: 62-71
  • DOI: 10.1016/j.biocel.2015.02.013
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Signaling pathway for adiponectin expression in adipocytes by osteocalcin. (2015)
  • Author(s): Otani, T., Mizokami, A., Hayashi, Y., Gao, J., Mori, Y., Nakamura, S., Takeuchi, H. and Hirata, M.
  • Journal Title: Cellular Signalling Volume: 27 Issue: 3 Pages: 532-544
  • DOI: 10.1016/j.cellsig.2014.12.018
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Hetero-oligomerization of C2 domains of phospholipase C-related but catalytically inactive protein and synaptotagmin-1. (2015)
  • Author(s): Wang, D., Takeuchi, H., Gao, J., Zhang, Z. and Hirata, M.
  • Journal Title: Advances in Biological Regulation Volume: 57 Pages: 120-129
  • DOI: 10.1016/j.jbior.2014.09.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Oral administration of osteocalcin improves glucose utilization by stimulating glucagon-like peptide-1 secretion. (2014)
  • Author(s): Mizokami, A., Yasutake, Y., Higashi, S., Kawakubo-Yasukochi, T., Chishaki, S., Takahashi, I., Takeuchi, H. and Hirata, M.
  • Journal Title: Bone Volume: 69 Pages: 68-79
  • DOI: 10.1016/j.bone.2014.09.006
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] PRIP (phospholipase C-related but catalytically inactive protein) inhibits exocytosis by direct interactions with syntaxin 1 and SNAP-25 through its C2 domain (2013)
  • Author(s): Zhang, Z.
  • Journal Title: The Journal of Biological Chemistry Volume: 288 Issue: 11 Pages: 7769-7780
  • DOI: 10.1074/jbc.m112.419317
  • Peer Reviewed
• [Journal Article] Phospholipase C-related catalytically inactive protein, a novel microtubule-associated protein 1 light chain 3-binding protein, negatively regulates autophagosome formation (2013)
  • Author(s): Umebayashi, H.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 432 Issue: 2 Pages: 268-274
  • DOI: 10.1016/j.bbrc.2013.01.119
  • Peer Reviewed
• [Journal Article] Osteocalcin induces release of glucagon-like peptide-1 and thereby stimulates insulin secretion in mice. (2013)
  • Author(s): Akiko Mizokami, Yu Yasutake, Jing Gao, Miho Matsuda, Ichiro Takahashi, Hiroshi Takeuchi, Masato Hirata
  • Journal Title: Plos ONE Volume: 8 Issue: 2 Pages: e57375-e57375
  • DOI: 10.1371/journal.pone.0057375
  • Peer Reviewed
• [Journal Article] Phospholipase C-related but catalytically inactive protein, PRIP as a scaffolding protein for phospho-regulation (2013)
  • Author(s): Sugiyama et al
  • Journal Title: Advances in Bioloical Regulation Volume: 53 Issue: 3 Pages: 331-340
  • DOI: 10.1016/j.jbior.2013.07.001
  • Peer Reviewed
• [Journal Article] Regulated interaction of protein phosphatase 1 and protein phosphatase 2A with phospholipase C-related but catalytically inactive protein. (2012)
  • Author(s): Goro Sugiyama, Hiroshi Takeuchi, Koki Nagano, Jing Gao, Yukiko Ohyama, Yoshide Mori, Masato Hirata
  • Journal Title: Biochemistry Volume: 51 Issue: 16 Pages: 3394-403
  • DOI: 10.1021/bi2018128
  • Peer Reviewed
• [Journal Article] Phospholipase C-related but catalytically inactive protein (PRIP) modulates synaptosomal-associated protein 25 (SNAP-25) phosphorylation and exocytosis. (2012)
  • Author(s): Jing Gao, Hiroshi Takeuchi, Zhao Zhang, Mitsunori Fukuda and Masato Hirata
  • Journal Title: The Journal of Biological Chemistry Volume: 287 Issue: 13 Pages: 10565-10578
  • DOI: 10.1074/jbc.m111.294645
  • Peer Reviewed
• [Presentation] Hetero-oligomerization of C2 domains of phospholipase C-related but catalytically inactive protein and synaptotagmin-1. (2015)
  • Author(s): Wang, D., Takeuchi, H., Gao, J., Zhang, Z., Hirata, M.
  • Organizer: Kyudai Oral Bioscience 2015 -9th International Symposium-
  • Place of Presentation: 福岡リーセントホテル（福岡県福岡市）
  • Year and Date: 2015-02-28
• [Presentation] Osteocalcin modifies insulin-induced glucose-uptake in cultured myotube. (2015)
  • Author(s): Tsuka, S., Aonuma, F., Hosokawa, R., Hirata, M., Takeuchi, H.
  • Organizer: APC2015
  • Place of Presentation: 九州歯科大学講堂（福岡県北九州市）
  • Year and Date: 2015-01-24
• [Presentation] 培養筋芽細胞を用いたオステオカルシンによる血糖調節修飾作用の検討 (2014)
  • Author(s): 柄慎太郎、青沼史子、細川隆司、東泉、大住伴子、平田雅人、竹内弘
  • Organizer: 第67回日本薬理学会西南部会
  • Place of Presentation: 産業医科大学（福岡県北九州市）
  • Year and Date: 2014-11-23
• [Presentation] Phosphorylation of SNAP-25 by protein kinase-A is negatively involved in calcium-dependent exocytosis in PC12 cells. (2014)
  • Author(s): Gao, J., Takeuchi, H., Wang D. and Hirata, M.
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Interaction of the C2 domains from different molecules. (2014)
  • Author(s): Wang, D., Takeuchi, H., Gao, J., Hirata, M.
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Involvement of tomosyn in insulin-dependent GLUT4 exocytosis as an Akt-substrate. (2014)
  • Author(s): Nagano, K., Takeuchi, H., Gao, J., Otani, T., Hirata, M.
  • Organizer: 39th Hormone and Cell Regulation
  • Place of Presentation: Mont Sainte Odile, France
  • Year and Date: 2014-10-09 – 2014-10-12
• [Presentation] 骨格筋に対するオステオカルシンの効果について， (2014)
  • Author(s): 柄慎太郎、青沼史子、細川隆司、平田雅人、竹内弘
  • Organizer: 第56回歯科基礎医学会学術大会
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] オステオカルシン受容体候補タンパク質 GPRC6A の免疫組織学的解析 (2014)
  • Author(s): 東泉、溝上顕子、大住伴子、平田雅人、竹内弘
  • Organizer: 第56回歯科基礎医学会学術大会
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] オステオカルシン経口投与時の体内動態と糖代謝への作用 (2014)
  • Author(s): 溝上顕子、安武雄、東泉、川久保-安河内友世、高橋一郎、竹内弘、平田雅人
  • Organizer: 第56回歯科基礎医学会学術大会
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Effect of osteocalcin on skeletal muscle cells. (2014)
  • Author(s): Tsuka, S., Aonuma, F., Hosokawa, R., Hirata, M. and Takeuchi, H.
  • Organizer: IADR General Session
  • Place of Presentation: Capetown, South Africa
  • Year and Date: 2014-06-25 – 2014-06-28
• [Presentation] Analysis of regulated interaction between Akt and PRIP. (2013)
  • Author(s): 竹内弘、杉山悟郎、長野公喜、大谷崇仁、高靖、平田雅人
  • Organizer: 第 86 回日本生化学会
  • Place of Presentation: 横浜市
• [Presentation] Akt による開口分泌調節タンパク質 tomosyn のリン酸化の検討 (2013)
  • Author(s): 長野公喜、竹内弘、高靖、杉山悟郎、大谷崇仁、平田雅人
  • Organizer: 第 86 回日本生化学会
  • Place of Presentation: 横浜市
• [Presentation] Differential role of SNAP-25 phosphorylation by protein kinase A and C in SNARE complex formation. (2013)
  • Author(s): Gao, J, Takeuchi, H., Wang, D., Hirata, M
  • Organizer: 第 86 回日本生化学会
  • Place of Presentation: 横浜市
• [Presentation] GLUT4 トランスロケーションにおける Akt による tomosyn のリン酸化の影響 (2013)
  • Author(s): 長野公喜、竹内弘、杉山悟郎、大谷崇仁、平田雅人
  • Organizer: 第 55 回歯科基礎医学会学術大会
  • Place of Presentation: 岡山市
• [Presentation] Molecular characterization of PRIP as an adaptor protein of Akt (2013)
  • Author(s): 杉山悟郎、竹内弘、長野公喜、大谷崇仁、平田雅人
  • Organizer: 第 55 回歯科基礎医学会学術大会
  • Place of Presentation: 岡山市
• [Presentation] Identification of tomosyn as a new downstream target of Akt. (2013)
  • Author(s): Nagano, K., Takeuchi, H., Sugiyama, G., Gao, J., Otani, T., Hirata, M.
  • Organizer: EB2013
  • Place of Presentation: Boston, MA, USA
• [Presentation] Phosphorylation-dependent interaction of PRIP with Akt. (2013)
  • Author(s): ugiyama, G., Nagano, K., Otani, T., Gao, J., Takeuchi, T., Hirata, M.
  • Organizer: EB2013
  • Place of Presentation: Boston, MA, USA
• [Presentation] Involvement of Phospholipase C-related but catalytically inactive protein in regulated exocytosis through the pleckstrin homology and the C2 domains. (2013)
  • Author(s): Takeuchi, H., Zhang, Z., Gao, J., Wang, DG., Hirata, M.
  • Organizer: EB2013
  • Place of Presentation: Boston, MA, USA
• [Presentation] Phosphorylation of tomosyn by Akt is implicated in the regulation of GLUT4 translocation. (2013)
  • Author(s): Nagano, K. Takeuchi, H., Gao, J., Otani, T., Hirata, M.
  • Organizer: The 8th Japan-Korea Conference on Celllular Signaling for Young Scientists
  • Place of Presentation: 福岡市
• [Presentation] Negative regulation of exocytosis by PRIP through intermolecular interactions. (2013)
  • Author(s): Takeuchi, H., Zhang, Z., Gao, J., Wang, D., Hirata, M.
  • Organizer: 第86回日本薬理学会年会
  • Place of Presentation: 福岡市
• [Presentation] Regulation of Phospholipase C-related but Catalytically Inactive Protein in Exocytosis by Direct Interaction with Syntaxin 1 and SNAP-25. (2013)
  • Author(s): Zhang, Z., Takeuchi, H., Gao, J., Wang, D., Hirata, M.
  • Organizer: International Symposium on Human Resource Deelopment towards Global Initiative
  • Place of Presentation: タイ王国
• [Presentation] Direct interaction of phospholipase C-related but catalytically inactive protein with syntaxin 1A and SNAP25 through C2 domain. (2012)
  • Author(s): Zhang, Z., Takeuchi, H., Gao, J., Wang, D., Hirata, M.
  • Organizer: 22nd IUBMB & 37th FEBS Congress
  • Place of Presentation: スペイン
• [Presentation] Role of phosphorylation of SNAP-25 by protein kinase A on SNARE complex formation. (2012)
  • Author(s): Gao, J., Takeuchi, H., Hirata, M.
  • Organizer: 22nd IUBMB & 37th FEBS Congress
  • Place of Presentation: スペイン
• [Remarks] 所属研究室ホームページ
  • URL: http://www.kyu-dent.ac.jp/research/lecture/oral_pharmacological
• [Remarks] 所属分野ホームページ
  • URL: http://www.kyu-dent.ac.jp/research/lecture/oral_pharmacological