シナプスタンパク質ユビキチン様修飾系の生理機能と神経疾患への関与

2011 Fiscal Year Annual Research Report

• Project/Area Number: 21500347
• Research Institution: Shinshu University
• Principal Investigator: 鈴木 龍雄 信州大学, 医学系研究科, 教授 (80162965)
• Co-Investigator(Kenkyū-buntansha): 棚橋 浩 信州大学, 医学系研究科, 准教授 (90236654)
• Keywords: PSD / シナプス後肥厚部 / シナプス / タンパク質修飾

Research Abstract

Ufm1修飾系(最も新しいUbiquitin様のタンパク質修飾系)の神経活動における役割を明らかにする為にUfm1基質の同定を試みた。2種のUfm1結合タンパク質(CDK5RAP3 [CDK5 regulatory subunit-associated protein 3], PRKABI [5'-AMP-activated protein kinase subunit beta-1]),4種のUfc1結合タンパク質(CDK5RAP3, Gstcd [glutathion S-transferase, C-terminal domain containing], MEF2C [myocyte-specific enhancer factor 2C], VHL [Von Hippel-Lindau disease tumor suppressor])にV5-Hisタグを付けた蛋白とFlag-Ufm1, Uba5, Ufc1をCOS7細胞で共発現させてUfm1の基質となり得るかを調べた。次に,COS7細胞にFlag-His-Ufm1-HA, Ufc1, Uba5を発現させ,その細胞ライセートから尿素可溶性画分を作成し,Ni-NTAカラムにかけてFlag-His-Ufm1化蛋白を精製し,Western blottingと質量分析法により数本の候補バンドを同定した。それらはUfm1化されたUba5,その分解物,Ufm1化されたUfc1の他に25kDaのバンドであった。この25kDaのバンドをLC-MS/MSにより解析すると,thbredoxin, RPL26L1 (ribosomal protein L26-like 1), Q6DRA9, histone H4が候補であった。これら蛋白にV5-Hisタグを付けたタンパク質とFlag-Ufm1, Uba5, Ufc1をCOS7細胞で共発現させてUfm1の基質となり得るかを調べた。

Research Products

• [Journal Article] SynArfGEF is a guanine nucleotide exchange factor for Arf6 and localizes preferentially at postsynaptic specializations of inhibitory synapses (2011)
  • Author(s): Fukaya M, Kamata A, Hara Y, Tamaki H, Katsumata O, Ito N, Takeda S, Hata Y, Suzuki T, Watanabe M, Harvey RJ and Sakagami H
  • Journal Title: J. Neurochem Volume: 116 Pages: 1122-1137
  • DOI: DOI:10.1111/j.1471-4159.2010.07167.x
  • Peer Reviewed
• [Journal Article] Association of Membrane Rafts and Postsynaptic Density : Proteomics, Biochemical, and Ultrastructural Analyses (2011)
  • Author(s): Suzuki T, Zhang J, Miyazawa S, Liu Q, Farzan MR, Yao WD
  • Journal Title: J.Neurochem. Volume: 119 Pages: 64-77
  • DOI: 10.1111/j.1471-4159.2011.07404.x
  • Peer Reviewed
• [Journal Article] タンパク質合成と可塑性 (2011)
  • Author(s): 鈴木龍雄
  • Journal Title: Clinical Neuroscience 29 Volume: 29 Pages: 763-765
• [Presentation] Association of postsynaptic density and membrane raft domainシナプス後肥厚部-メンブランラフト複合体の解析 (2011)
  • Author(s): Tatsuo Suzuki, Qian Liu
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2011-12-13
• [Presentation] Colocalization of Hippocampal Cholinergic Neurostimulating Peptide (HCNP) precursor protein with collapsin response mediator protein (CRMP)-2 at pre synapse in the hippocampus (2011)
  • Author(s): Kato D, Matsukawa N, Kanamori T, Mizuno M, Suzuki T, Ojika K
  • Organizer: Society for Neuroscience
  • Place of Presentation: Washington
  • Year and Date: 2011-11-13
• [Presentation] Analysis of association of PSD with postsynaptic membrane rafts (2011)
  • Author(s): Suzuki T, Liu Q
  • Organizer: Society for Neuroscience
  • Place of Presentation: Washington
  • Year and Date: 2011-11-12
• [Book] Isolation of synapse-subdomains by subcellular fractionation using sucrose density gradient centrifugation. in Neuroproteomics in the series "Neuromethods" (Springer Protocols) (2011)
  • Author(s): Tatsuo Suzuki
  • Total Pages: 15(317)
  • Publisher: Humana Press