Determination of PKC substrate involved in neurodegenerative diseases and its application for drug design

• Project/Area Number: 25293060
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: General pharmacology
• Research Institution: Kobe University
• Principal Investigator: SAITO NAOAKI 神戸大学, バイオシグナル研究センター, 教授 (60178499)
• Co-Investigator(Kenkyū-buntansha): ADACHI Naoko 神戸大学, バイオシグナル研究センター, 助教 (70604510) ; UEYAMA Takehiko 神戸大学, バイオシグナル研究センター, 准教授 (80346254)
• Co-Investigator(Renkei-kenkyūsha): SHIRAFUJI Toshihiko 神戸大学, バイオシグナル研究センター, 研究員 (30595765)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000); Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2013: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: リン酸化酵素 / パーキンソン病 / モデル動物 / 治療薬 / リン酸化プロテオーム / プロテインキナーゼC / モデルマウス / 質量分析 / ドーパミン

Outline of Final Research Achievements

We identified substrate proteins for PKCg as a new drug target for Parkinson disease. Nine substrate proteins were identified: Connexin-43, Disk1, MADD, CSPa, Calnexin, Stathmin, bPIX, NogoA, Adducin. Furthermore, 1) Phosphorylation of Ser583, Ser340 of bPIX is involved in DA release, 2) Phosphorylation of Connexin-43, MADD, Calnexin and Adducin is also related to DA release, 3) Phosphorylation of Ser10 of CSPa is important for cell viability.

Research Products

• [Journal Article] Essential role of constitutive androstane receptor in Ginkgo biloba extract induced liver hypertrophy and hepatocarcinogenesis (2015)
  • Author(s): Maeda J, Inoue K, Ichimura R, Takahashi M, Kodama Y, Saito N, Yoshida M.
  • Journal Title: Food Chem Toxicol. Volume: 83 Pages: 201-209
  • DOI: 10.1016/j.fct.2015.06.010
  • Peer Reviewed / Open Access
• [Journal Article] The Extracellular A-loop of Dual Oxidases Affects the Specificity of Reactive Oxygen Species Release. (2015)
  • Author(s): Ueyama T, Sakuma M, Ninoyu Y, Hamada T, Dupuy C, Geiszt M, Leto TL, Saito N.
  • Journal Title: J Biol. Chem Volume: 290 Issue: 10 Pages: 6495-6506
  • DOI: 10.1074/jbc.m114.592717
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification and characterization of PKCγ, a kinase associated with SCA14, as an amyloidogenic protein. (2015)
  • Author(s): Takahashi H, Adachi N, Shirafuji T, Danno S, Ueyama T, Vendruscolo M, Shuvaev AN, Sugimoto T, Seki T, Hamada D, Irie K, Hirai H, Sakai N, *Saito N.
  • Journal Title: Hum Mol Genet. Volume: 24 Issue: 2 Pages: 525-539
  • DOI: 10.1093/hmg/ddu472
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The role of Pak-interacting exchange factor-β phosphorylation at serines 340 and 583 by PKCγ in dopamine release. (2014)
  • Author(s): Shirafuji T, Ueyama T, Yoshino K, Takahashi H, Adachi N, Ago Y, Koda K, Nashida T, Hiramatsu N, Matsuda T, Toda T, Sakai N, Saito N.
  • Journal Title: J Neurosci Volume: 34 Issue: 28 Pages: 9268-9280
  • DOI: 10.1523/jneurosci.4278-13.2014
  • NAID: 120005619641
  • Peer Reviewed / Open Access
• [Journal Article] Maintenance of stereocilia and apical junctional complexes by Cdc42 in cochlear hair cells (2014)
  • Author(s): Ueyama T, Sakaguchi H, Nakamura T, Goto A, Morioka S, Shimizu A, Nakao K, Hishikawa Y, Ninoyu Y, Kassai H, Suetsugu S, Koji T, Fritzsch B, Yonemura S, Hisa Y, Matsuda M, Aiba A, Saito N.
  • Journal Title: J Cell Sci. Volume: 127 Pages: 2040-2052
  • DOI: 10.1242/jcs.143602
  • NAID: 120006986582
  • Peer Reviewed / Open Access
• [Journal Article] Heparin inhibits melanosome uptake and inflammatory response coupled with phagocytosis through blocking PI3k/Akt and MEK/ERK signaling pathways in human epidermal keratinocytes. (2014)
  • Author(s): Makino-Okamura C, Niki Y, Takeuchi S, Nishigori C, Declercq L, Yaroch DB, Saito N.
  • Journal Title: Pigment Cell Melanoma Res. Volume: 27 Issue: 6 Pages: 1063-1074
  • DOI: 10.1111/pcmr.12287
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] RANTES/CCL5 mediated-biological effects depend on the syndecan-4/PKCa signaling pathway (2014)
  • Author(s): Maillard, L., Saito, N., Hlawaty, H., Friand, V., Suffee, N., Chmilewsky, F., Haddad, O., Laguillier, C., Guyot, E., Ueyama, T., Oudar, O., Sutton, A., Charnaux, N.
  • Journal Title: Biology Open Volume: 3 Issue: 10 Pages: 995-1004
  • DOI: 10.1242/bio.20148227
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Both the C1 domain and a basic amino acid cluster at C-terminus are important for neurite and branch induction ability of DGK. (2014)
  • Author(s): Takuya Kano, Takeshi Kouzuki, Satoru Mizuno, Shuji Ueda, Minoru Yamanoue, Fumio Sakane, Naoaki Saito and Yasuhito Shirai
  • Journal Title: iochem. Biophys. Res. Commun Volume: 印刷中 Issue: 1 Pages: 89-94
  • DOI: 10.1016/j.bbrc.2014.03.113
  • Peer Reviewed / Open Access
• [Journal Article] Diacylglycerol kinase gamma regulates antigen-induced mast cell degranulation by mediating Ca2+ influxes (2014)
  • Author(s): Sakuma, M., Shirai, Y., Ueyama, T., and Saito, N.
  • Journal Title: Biochem. Biophys. Res. Commun, Volume: 445 Issue: 2 Pages: 340-345
  • DOI: 10.1016/j.bbrc.2014.01.197
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Deregulation of the actin cytoskeleton and macropinocytosis in response to phorbol ester by the mutant protein kinase C gamma that causes spinocerebellar ataxia type 14. (2014)
  • Author(s): Yamamoto K, Seki T, Yamamoto H, Adachi N, Tanaka S, Hide I, Saito N, Sakai N
  • Journal Title: Front Physiol. Volume: 5 Pages: 126-126
  • DOI: 10.3389/fphys.2014.00126
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Diacylglycerol kinase as a possible therapeutic target for neuronal diseases. (2014)
  • Author(s): Shirai, Y and Saito, N
  • Journal Title: Journal of Biomedical Science Volume: 21 Pages: 28-36
  • Peer Reviewed
• [Journal Article] Maintenance of stereocilia and apical junctional complexes by Cdc42 in cochlear hair cells, (2014)
  • Author(s): Ueyama, T., Sakaguchi, H., Nakamura, T., Goto, A., Morioka, S., Shimizu, A., Nakao, K., Hishikawa, Y., Ninoyu, Y., Kassai, H., Suetsug, J., Koji, T., Fritzsch, B., Yonemura, S., Hisa, Y., Matsuda, M., Aiba, A., and Saito, N.
  • Journal Title: J. Cell Science Volume: 印刷中
  • NAID: 120006986582
  • Peer Reviewed
• [Journal Article] Negative Charges in the Flexible N-Terminal Domain of Rho GDP-Dissociation Inhibitors (RhoGDIs) Regulate the Targeting of the RhoGDI-Rac1 Complex to Membranes (2013)
  • Author(s): Ueyama T, Son J, Kobayashi T, Hamada T, Nakamura T, Sakaguchi H, Shirafuji T, Saito N
  • Journal Title: J. Immunol. Volume: 191 Issue: 5 Pages: 2560-2569
  • DOI: 10.4049/jimmunol.1300209
  • Peer Reviewed
• [Journal Article] Mutant gammaPKC that causes spinocerebellar ataxia type 14 upregulates Hsp70, which protects cells from the mutant's cytotoxicity. (2013)
  • Author(s): Ogawa, K., Seki, T., Onji, T., Adachi, N., Tanaka, S., Hide, I., Saito, N., Sakai, N.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 440 Issue: 1 Pages: 25-30
  • DOI: 10.1016/j.bbrc.2013.09.013
  • Peer Reviewed
• [Journal Article] PKC-ε pseudosubstrate and catalytic activity are necessary for membrane delivery during IgG-mediated phagocytosis. (2013)
  • Author(s): Wood, TR., Chow, RY., Hanes, CM., Zhang, X., Kashiwagi, K., Shirai, Y., Trebak, M., Loegering, DJ., Saito, N., and Lennartz, MR.
  • Journal Title: J Leukoc Biol Volume: 94 Issue: 1 Pages: 109-122
  • DOI: 10.1189/jlb.1212634
  • Peer Reviewed
• [Journal Article] Increased seizure susceptibility in a mouse with diacylglycerol kinase β 2 deficiency (2013)
  • Author(s): Ishisaka, M., Tsuruma, K., Shimazawa, M., Shirai, Y., Saito, N., and Hara, H.
  • Journal Title: Neuroscience & Medicine Volume: 4 Pages: 117-122
  • Peer Reviewed
• [Journal Article] Protein kinase Cη is targeted to lipid droplets. (2013)
  • Author(s): Suzuki Michitaka
  • Journal Title: Histochem Cell Biol Volume: 139 Issue: 4 Pages: 505-511
  • DOI: 10.1007/s00418-013-1083-z
  • Peer Reviewed
• [Journal Article] Both C1B domain and pseudosubstrate region are necessary for saturated fatty acid membrane;distinct role of intramolecular domains for different (2013)
  • Author(s): Takaaki Nishimoto, T., Kashiwagi, K., Saito, N., and Shirai, Y.
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 432 Issue: 2 Pages: 384-388
  • DOI: 10.1016/j.bbrc.2013.01.048
  • Peer Reviewed
• [Presentation] Protein kinase C and its involvement in neuronal diseases (2016)
  • Author(s): Naoaki Saito
  • Organizer: Neuroscience Network in Kobe シンポジウム
  • Place of Presentation: 神戸大学（兵庫県・神戸市）
  • Year and Date: 2016-02-19
  • Invited
• [Presentation] 脊髄小脳変性症14型におけるPKCのアミロイド様構造体の形成と疾患への関与 (2015)
  • Author(s): 足立直子、高橋英之、白藤俊彦、上山健彦、Michele, Vendruscolo., 入江一浩、平井宏和、酒井規雄、齋藤尚亮
  • Organizer: 第38回日本神経科学大会
  • Place of Presentation: 神戸国際会議場（兵庫県・神戸市）
  • Year and Date: 2015-07-28
• [Presentation] Cdc42 is required for maintenance of stereocilia and apical junctional complexes in cochlear hair cells. (2014)
  • Author(s): Ueyama, T., Sakaguchi, H., Goto, A., Hisa, Y., Matsuda, M., Aiba, A. and Saito, N.
  • Organizer: The 37th Naito Conference
  • Place of Presentation: 北海道ニセコ
  • Year and Date: 2014-07-15 – 2014-07-18
• [Presentation] Cde-42 is required for maintenance of stereocilia and apical junctional complexes in cochlear hair cells (2014)
  • Author(s): Ueyama, T. and Saito, N.
  • Organizer: Joint Workshop between University Catholic Louvain and Kobe University
  • Place of Presentation: Brussels, Belgium
• [Presentation] A protein kinase Cgamma knockout mice as a Parkinsonian model: Involvement of Pak -Interacting Exchange Factorbeta (2014)
  • Author(s): 齋藤尚亮
  • Organizer: 第87回日本薬理学会年会
  • Place of Presentation: 仙台
• [Presentation] Protein kinase C and its involvement in neurodegenerative disease (2014)
  • Author(s): Naoaki Saito
  • Organizer: Symposium on Membrane Biology
  • Place of Presentation: Seattle WA, USA
  • Invited
• [Presentation] Both C1 domain and basic amino acid cluster at C-terminus are important for branching and neurite induction of DGK (2013)
  • Author(s): Shirai,Y., Takuya,K., Kouzuki,T., Ueda,S., Yamanoue,M., Sakane,F. and Saito,N.
  • Organizer: Neuro 2013
  • Place of Presentation: 京都
• [Presentation] 黒質線条体系におけるPKCγの基質の解析：β2PIXリン酸化のドパミン遊離への 役割 (2013)
  • Author(s): 白藤 俊彦、上山 健彦、吉野 健一、足立 直子、高橋 英之、香田 健、 平松 直樹、吾郷 由希夫、松田 敏夫、酒井 規雄、齋藤 尚亮
  • Organizer: 第124回日本薬理学会近畿部会
  • Place of Presentation: 京都