糖タンパク質選別輸送装置の超分子形成および作動機構の解明

• Project Area: Structural basis of cell-signalling complexes mediating signal perception, transduction and responses
• Project/Area Number: 25121730
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Nagoya City University
• Principal Investigator: 佐藤 匡史 名古屋市立大学, 大学院薬学研究科, 准教授 (80532100)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2013: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 構造生物学 / 積荷受容体 / 糖タンパク質 / X線結晶構造解析 / NMR解析

Outline of Annual Research Achievements

分泌経路の入り口である小胞体では、糖鎖を目印とした品質管理システムを通じて、タンパク質の立体構造形成、分解、および選別輸送が行われている。例えば、モノグルコシル化された糖鎖を有する糖タンパク質は立体構造形成の途上にあることを示唆している。
小胞体-ゴルジ体間における糖タンパク質の選別輸送は、細胞内レクチンERGIC-53による糖鎖認識と、カルシウム結合タンパク質MCFD2によるポリペプチド鎖認識が協働することにより担われている。本年度は、ERGIC53－MCFD2複合体の作動メカニズムを解明するために、MCFD2によるポリペプチド鎖認識の構造基盤を明らかにすることを目指した。そこでNMRおよび等温カロリメトリーによる相互作用解析を実施し、ERGIC53－MCFD2複合体による血液凝固因子のポリペプチド鎖認識の構造基盤に関する情報を収集した。
さらに本研究では、立体構造が未完成の糖タンパク質に選択的にモノグルコシル化するフォールディングセンサー酵素UGGTの基質認識ドメインの一部に関して3次元構造情報を得ることに初めて成功した。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Conformational dynamics of oligosaccharides characterized by paramagnetism-assisted NMR spectroscopy in conjunction with molecular dynamics simulation (2015)
  • Author(s): Ying Zhang
  • Journal Title: Adv. Exp. Med. Biol. Volume: 842 Pages: 217-230
  • DOI: 10.1007/978-3-319-11280-0_14
  • ISBN: 9783319112794, 9783319112800
  • Peer Reviewed
• [Journal Article] Emerging structural insights into glycoprotein quality control coupled with N-glycan processing in the endoplasmic reticulum (2015)
  • Author(s): T.Satoh, T.Yamaguchi, and K.Kato
  • Journal Title: Molecules Volume: 20 Issue: 2 Pages: 2475-2491
  • DOI: 10.3390/molecules20022475
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Recent advances in glycoprotein production for structural biology: toward tailored design of glycoforms (2014)
  • Author(s): Y.Kamiya, T.Satoh, and K.Kato
  • Journal Title: Curr. Opin. Struct. Biol. Volume: 26 Pages: 44-53
  • DOI: 10.1016/j.sbi.2014.03.008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Crystal structure of archaeal homolog of proteasome-assembly chaperone PbaA (2014)
  • Author(s): Arunima Sikar
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 453 Issue: 3 Pages: 493-497
  • DOI: 10.1016/j.bbrc.2014.09.114
  • Peer Reviewed
• [Journal Article] Mode of substrate recognition by the Josephin domain of ataxin-3, which has an endo-type deubiquitinase activity (2014)
  • Author(s): T.Satoh, A.Sumiyoshi, M.Yagi-Utsumi, E.Sakata, H.Sasakawa, E.Kurimoto, Y.Yamaguchi, W.Li, C.A.P. Joazeiro, T.Hirokawa, and K.Kato
  • Journal Title: FEBS Lett. Volume: 588 Issue: 23 Pages: 4422-4430
  • DOI: 10.1016/j.febslet.2014.10.013
  • Peer Reviewed
• [Journal Article] Structural insight into substrate recognition by the endoplasmic reticulum folding-sensor enzyme: crystal structure of third thioredoxin-like domain of UDP-glucose:glycoprotein glucosyltransferase (2014)
  • Author(s): T.Zhu, T.Satoh, and K.Kato
  • Journal Title: Sci. Rep. Volume: 4 Issue: 1 Pages: 7322-7322
  • DOI: 10.1038/srep07322
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Structural basis for disparate sugar-binding specificities in the homologous cargo receptors ERGIC-53 and VIP36 (2014)
  • Author(s): Tadashi Satoh, Kousuke Suzuki, Takumi Yamaguchi, and Koichi Kato
  • Journal Title: PLoS ONE Volume: 9 Issue: 2 Pages: e87963-e87963
  • DOI: 10.1371/journal.pone.0087963
  • Peer Reviewed
• [Journal Article] ATPase activity and ATP-dependent conformational change in the co-chaperone HOP. (2014)
  • Author(s): Yamamoto S, Subedi GP, Hanashima S, Satoh T, Otaka M, Wakui H, Sawada KI, Yokota SI, Yamaguchi Y, Kubota H, Itoh H.
  • Journal Title: Journal of Biological Chemistry Volume: 289 Issue: 14 Pages: 9880-9886
  • DOI: 10.1074/jbc.m114.553255
  • Peer Reviewed
• [Journal Article] Structural Basis for Proteasome Formation Controlled by an Assembly Chaperone Nas2 (2014)
  • Author(s): Satoh T, Saeki Y, Hiromoto T, Wang YH, Uekusa Y, Yagi H, Yoshihara H, Yagi-Utsumi M, Mizushima T, Tanaka K, Kato K
  • Journal Title: Structure Volume: (掲載確定) Issue: 5 Pages: 731-743
  • DOI: 10.1016/j.str.2014.02.014
  • Peer Reviewed
• [Journal Article] Recognition of Bisecting N-Acetylglucosamine: Structural basis for asymmetric interaction with the mouse lectin dendritic cell inhibitory receptor 2 (2013)
  • Author(s): Masamichi Nagae
  • Journal Title: J. Biol. Chem. Volume: 288 Issue: 47 Pages: 33598-33610
  • DOI: 10.1074/jbc.m113.513572
  • Peer Reviewed
• [Presentation] Structural basis for sugar trimming reactions by glucosidase II involved glycoprotein quality control system (2015)
  • Author(s): Tadashi Satoh
  • Organizer: The 5th Yonsei-IMS Joint Workshop: “Multilateral approaches for investigating biomolecular structures and functions”
  • Place of Presentation: 箱根
  • Year and Date: 2015-01-31
• [Presentation] Structural aspects of glycoprotein folding sensor enzyme (UGGT): A combined X-ray crystallography and small angle X-ray scattering study (2015)
  • Author(s): Tong Zhu
  • Organizer: The 5th Yonsei-IMS Joint Workshop: “Multilateral approaches for investigating biomolecular structures and functions”
  • Place of Presentation: 箱根
  • Year and Date: 2015-01-31
• [Presentation] Structural basis for recognition of the terminal glucose tag of N-glycans as fate-determinant of glycoproteins in cells (2014)
  • Author(s): Koichi Kato
  • Organizer: The 7th Korea-Japan Seminars on Biomolecular Sciences: Experiments and Simulations
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2014-11-26
  • Invited
• [Presentation] UDP-glucose: glycoprotein glucosyltransferase (UGGT) possesses three-tandem thioredoxin-like domains (2014)
  • Author(s): Tong Zhu
  • Organizer: 第87回 日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-18
• [Presentation] Structural insights into glycoprotein processing mechanisms in ER quality control system (2014)
  • Author(s): 佐藤匡史
  • Organizer: 第87回 日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15
  • Invited
• [Presentation] Structural basis for the glycan-processing mechanisms in glycoprotein quality control system (2014)
  • Author(s): 佐藤匡史
  • Organizer: 第52回 日本生物物理学会年会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-26
  • Invited
• [Presentation] 小胞体グルコシダーゼIIの基質認識メカニズムの構造基盤 (2014)
  • Author(s): 年森隆泰
  • Organizer: 第33回 日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-11
• [Presentation] Structural basis for substrate recognition mechanism of ER glucosidase II (2014)
  • Author(s): Tadashi Satoh
  • Organizer: 23rd Congress and General Assembly of the International Union of Crystallography (IUCr 2014)
  • Place of Presentation: Montreal, Canada
  • Year and Date: 2014-08-07
• [Presentation] 糖タンパク質の細胞内プロセッシングに関わるグルコシダーゼⅡの基質認識メカニズム (2014)
  • Author(s): 年森隆泰
  • Organizer: 第12回 次世代を担う若手のためのフィジカル・ファーマフォーラム
  • Place of Presentation: 箱根
  • Year and Date: 2014-07-14
• [Presentation] 小胞体品質管理システムに関わるグルコシダーゼIIの基質認識機構 (2014)
  • Author(s): 佐藤 匡史
  • Organizer: 第14回 日本蛋白質科学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-06-26
• [Presentation] 糖タンパク質選別輸送装置の超分子形成および作動機構の解明 (2014)
  • Author(s): 佐藤 匡史
  • Organizer: 平成26年度新学術領域「構造細胞生物学」全体会議・総括班会議
  • Place of Presentation: ルスツ、北海道
  • Year and Date: 2014-06-15
• [Presentation] カーゴレセプターERGIC-53/MCFD2複合体による糖タンパク質認識メカニズムの構造基盤 (2014)
  • Author(s): 鈴木康介
  • Organizer: 第78回 日本生化学会中部支部例会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-24
• [Presentation] 糖タンパク質品質管理に関わるグルコシダーゼIIの基質認識メカニズムの構造基盤 (2014)
  • Author(s): 年森隆泰
  • Organizer: 第78回 日本生化学会中部支部例会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-24
• [Presentation] Structural insights into substrate recognition mechanism of glycoprotein processing enzyme ER glucosidase II (2014)
  • Author(s): Tadashi Satoh
  • Organizer: The 4th Asia Pacific Protein Association (APPA) Conference
  • Place of Presentation: Jeju, Korea
  • Year and Date: 2014-05-18
• [Presentation] Structural insights into proteasome orchestration mechanism through a proteasome assembly factor Nas2
  • Author(s): Tadashi Satoh
  • Organizer: 8th Asian Biophysics Association Symposium (ABA Jeju 2013)
  • Place of Presentation: Jeju, Korea
• [Presentation] 糖タンパク質選別輸送装置の超分子形成および作動機構の解明
  • Author(s): 佐藤匡史
  • Organizer: 平成25年度 新学術領域「構造細胞生物学」全体会議・総括班会議
  • Place of Presentation: 清里
• [Presentation] ERGIC-53/MCFD2 複合体による糖鎖認識の構造基盤
  • Author(s): 佐藤匡史
  • Organizer: 第59回 日本薬学会東海支部会
  • Place of Presentation: 名古屋
• [Presentation] Mechanistic insights into the proteasome orchestration by a proteasome assembly chaperone Nas2
  • Author(s): Tadashi Satoh
  • Organizer: 7th International Conference on Structural Genomics (ICSG2013-SLS)
  • Place of Presentation: 札幌
• [Presentation] Structural basis underlying the redox-dependent domain rearrangement of the substrate-binding region of protein disulfide isomerase
  • Author(s): Koya Inagaki
  • Organizer: 7th International Conference on Structural Genomics (ICSG2013-SLS)
  • Place of Presentation: 札幌
• [Presentation] 積荷受容体ERGIC-53/MCFD2による糖鎖認識の構造基盤
  • Author(s): 佐藤匡史
  • Organizer: 第32回日本糖質学会年会
  • Place of Presentation: 大阪
• [Presentation] Bacterial expression and EM analysis of UDP-glucose glycosyltransferase from a thermophilic fungus
  • Author(s): Tong Zhu
  • Organizer: 糖鎖科学中部拠点 第11回若手の力フォーラム
  • Place of Presentation: 名古屋
• [Presentation] EM tomography analysis of ER folding sensor enzyme UDP-glucose: glycoprotein glucosyltransferase
  • Author(s): Tong Zhu
  • Organizer: 平成25年度 生理学研究所研究会「電子顕微鏡機能イメージングの医学・生物学への応用」
  • Place of Presentation: 岡崎
• [Presentation] Interaction mode of a proteasome assembly chaperone Nas2 with the proteasome Rpt5 subunit
  • Author(s): Ying-Hui Wang
  • Organizer: 山田研究会・統合バイオサイエンスシンポジウム
  • Place of Presentation: 伊良湖
• [Presentation] Overall structure of ER folding sensor UDP-glucose: glycoprotein glucosyltransferase revealed by EM tomography
  • Author(s): Tong Zhu
  • Organizer: 山田研究会・統合バイオサイエンスシンポジウム
  • Place of Presentation: 伊良湖
• [Presentation] Redox-dependent conformational changes of the substrate-binding region of protein disulfide isomerase
  • Author(s): Koya Inagaki
  • Organizer: 山田研究会・統合バイオサイエンスシンポジウム
  • Place of Presentation: 伊良湖
• [Presentation] Structural basis for proteasome formation mediated by assembly chaperone Nas2
  • Author(s): Tadashi Satoh
  • Organizer: 6th Japan-Korea Seminars on Biomolecular Science : Experiments and Simulation
  • Place of Presentation: 岡崎
• [Presentation] An archaeal homolog of proteasome assembly chaperone forms a homotetramer and functions as proteasome activator
  • Author(s): Kentaro Kumoi
  • Organizer: 6th Japan-Korea Seminars on Biomolecular Science : Experiments and Simulation
  • Place of Presentation: 岡崎
• [Presentation] NMR characterization of the interaction of a proteasome assembly chaperone Nas2 with the proteasome Rpt5 subunit
  • Author(s): Ying-Hui Wang
  • Organizer: SOKENDAI Asian Winter School "Innovations and Challenges in Molecular Science: From Basics to Cutting-edge Researches”
  • Place of Presentation: 岡崎
• [Presentation] Structural basis for disparate sugar-binding specificities in the homologous cargo receptors ERGIC-53 and VIP36
  • Author(s): Tadashi Satoh
  • Organizer: Yonsei-IMS Seminars on Biomolecular Sciences : Protein Structures and Diseases
  • Place of Presentation: Busan, Korea
• [Presentation] The structural biology study on ER folding sensor UDP-glucose:glycoprotein glucosyltransferase (UGGT)
  • Author(s): Tong Zhu
  • Organizer: Yonsei-IMS Seminars on Biomolecular Sciences : Protein Structures and Diseases
  • Place of Presentation: Busan, Korea
• [Presentation] Structural-architecture of the ER glycoprotein folding sensor UDP-glucose:glycoprotein glucosyltransferase (UGGT)
  • Author(s): Tong Zhu
  • Organizer: 2013年度 生物物理中部支部会
  • Place of Presentation: 岡崎
• [Presentation] 小胞体品質管理に関わるグルコシダーゼIIの基質認識機構の構造基盤
  • Author(s): 年森隆泰
  • Organizer: 2013年度 生物物理中部支部会
  • Place of Presentation: 岡崎
• [Presentation] プロテインジスルフィドイソメラーゼの機能発現におけるミクロ‐マクロ相関の探査
  • Author(s): 稲垣宏弥
  • Organizer: 2013年度 生物物理中部支部会
  • Place of Presentation: 岡崎
• [Presentation] 小胞体糖タンパク質フォールディング装置を構成するタンパク質群の構造生物学研究
  • Author(s): 佐藤匡史
  • Organizer: 日本薬学会134年会
  • Place of Presentation: 熊本