Development of an in situ visible cell-free assay system for neurotransmitter release

• Project/Area Number: 25640036
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Kobe University
• Principal Investigator: SAKISAKA TOSHIAKI 神戸大学, 医学(系)研究科(研究院), 教授 (80359843)
• Co-Investigator(Kenkyū-buntansha): YAMAMOTO YASUNORI 神戸大学, 大学院医学研究科, 准教授 (30467659)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 神経伝達物質 / シナプス小胞 / 膜融合

Outline of Final Research Achievements

The main process of synaptic transmission form one neuron to another neuron is mediated by neurotransmitter release. The entry of Ca2+ into nerve terminals initiates the series of molecular events that culminates with fusion of the synaptic vesicle and release of neurotransmitter into the synaptic cleft. The presynaptic membrane curvature change has been shown to be involved in the synaptic vesicle fusion. Here we identify membrane-shaping proteins, Arl6IP1 and TMCC3. We also identify an Arl6IP1-interacting protein TMEM33. TMEM33 suppresses the membrane-shaping activity of Arl6IP1. On the other hand, TMCC3 has membrane-laminating activity through interacting with membrane-shaping proteins, reticulon family proteins.

Research Products

• [Journal Article] The Emerging Role of Calcium-modulating Cyclophilin Ligand (CAML) in Posttranslational Insertion of Tail-anchored Proteins into the Endoplasmic Reticulum Membrane. (2015)
  • Author(s): Yamamoto, Y., and Sakisaka, T.
  • Journal Title: The Journal of Biochemistry Volume: 印刷中 Issue: 6 Pages: 419-429
  • DOI: 10.1093/jb/mvv035
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Crystal structure of afadin PDZ domain-nectin-3 complex shows the structural plasticity of the ligand-binding site. (2015)
  • Author(s): Fujiwara, Y., Goda, N., Tamashiro, T., Narita, H., Satomura, K., Tenno, T., Nakagawa, A., Oda, M., Suzuki, M., Sakisaka, T., Takai, Y., and Hiroaki, H.
  • Journal Title: Protein Science Volume: 24 Pages: 376-385
  • Peer Reviewed
• [Journal Article] Cytoarchitecture of the olfactory bulb in the laggard mutant mouse. (2014)
  • Author(s): Yunus J, Setsu T, Kikkawa S, Sakisaka T, Terashima T.
  • Journal Title: Neuroscience Volume: 275 Pages: 259-271
  • DOI: 10.1016/j.neuroscience.2014.06.011
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification and Characterization of TMEM33 as a Reticulon-binding Protein. (2014)
  • Author(s): Urade, T., Yamamoto, Y., Zhang, X., Ku, Y., and Sakisaka, T.
  • Journal Title: Kobe Journal of Medical Sciences Volume: 60
  • NAID: 110009842833
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] VAP-B Binds to Rab3GAP1 at the ER: Its Implication in Nuclear Envelope Formation through the ER-Golgi Intermediate Compartment. (2014)
  • Author(s): Hantan, D., Yamamoto, Y., and Sakisaka, T.
  • Journal Title: Kobe Journal of Medical Sciences Volume: 60
  • NAID: 110009831834
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Arl6IP1 has the ability to shape the mammalian ER membrane in a reticulon-like fashion. (2014)
  • Author(s): Yamamoto, Y., Yoshida, A., Miyazaki, N., Iwasaki, K., and Sakisaka, T.
  • Journal Title: Biochemical Journal Volume: 458 Issue: 1 Pages: 69-79
  • DOI: 10.1042/bj20131186
  • Peer Reviewed
• [Presentation] 髄鞘低形成と皮質形成異常を示す新規突然変異マウスlaggard (2016)
  • Author(s): 匂坂 敏朗、寺島 俊雄
  • Organizer: 第121回日本解剖学会総会・全国学術集会
  • Place of Presentation: ビッグパレット福島
  • Year and Date: 2016-03-29
• [Presentation] 小胞体の膜形態制御と翻訳後膜挿入の分子機構 (2014)
  • Author(s): 山本 泰憲、匂坂 敏朗
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
  • Year and Date: 2014-10-15
• [Presentation] Reticulon結合タンパク質TMEM33の同定と性状解析 (2013)
  • Author(s): 張 霞, 山本 泰憲, 浦出 剛史, 匂坂 敏朗
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸国際会議場
• [Book] DOJIN BIOSCIENCEシリーズ『メンブレントラフィック』 PartIII:高次生体機能を支えるメンブレントラフィック １０章 神経伝達物質放出を支えるメンブレントラフィック (2016)
  • Author(s): 山本 泰憲、匂坂 敏朗
  • Publisher: 化学同人
• [Book] Presynaptic Terminals (2015)
  • Author(s): Yamamoto, Y., and Sakisaka, T.
  • Total Pages: 365
  • Publisher: Springer
• [Remarks]
  • URL: http://www.med.kobe-u.ac.jp/membrd/
• [Remarks] 神戸大学大学院医学研究科 生理学・細胞生物学講座 膜動態学分野
  • URL: http://www.med.kobe-u.ac.jp/membrd/
• [Remarks] 神戸大学大学院医学研究科 生理学・細胞生物学講座 膜動態学分野
  • URL: http://www.med.kobe-u.ac.jp/membrd/