細胞質Ngly1（PNGase）の中枢神経系における機能の解析

• Project Area: Deciphering sugar chain-based signals regulating integrative neuronal functions
• Project/Area Number: 26110725
• 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: Institute of Physical and Chemical Research
• Principal Investigator: 鈴木 匡 国立研究開発法人理化学研究所, 糖鎖代謝学研究チーム, チームリーダー (90345265)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2015: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
• Keywords: NGLY1 / 遺伝病 / 中枢神経系 / マウス / 行動解析 / Ngly1 / ENGase / 中枢神経 / 糖タンパク質 / 品質管理

Outline of Annual Research Achievements

Ngly1はタンパク質からN型糖鎖を切り離す酵素である。真核細胞に広く保存されている本酵素は、小胞体で発生した異常糖タンパク質がプロテアソームで分解される機構（小胞体関連分解(ERAD)）に関与している。2012年にNgly1遺伝子に欠損を持つヒト遺伝病の疾患が報告された。その症状は全身性であるが、運動障害や精神遅滞など、中枢神経系の異常によると疑われる症状が多く観察される。そこで本研究では中枢神経系におけるNgly1タンパク質の機能を明らかにするために、神経系で特異的に遺伝子がノックアウトされるNestin-Cre KOマウスを用いて、神経特異的Ngly1-KOマウスの表現型解析を行った。全身KOマウスは胚性後期-生後すぐに致死性を示すのと対照的に、神経特異的KOマウスは生存可能で、予想よりも緩和な表現型を示した。全身のNgly1 EngaseダブルKOマウスと比較しても後肢の反射異常はマイルドであった。組織切片の詳細な観察により、プルキンエ細胞の一部脱落、樹状突起の形態の軽微な異常等が観察されたが、行動解析においては顕著な異常は観察されなかった。一方で後ろ足で立つ動作（Rearing）の減少が見られたことから、中枢神経系のKOでは後肢にもっとも顕著な表現型が観察された。これらの結果から、NGLY1欠損症でみられる中枢神経系の症状は、別の組織の異常からくる２次的な影響である可能性が示唆された。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] The cytoplasmic peptide:N-glycanase (NGLY1); structure, expression and cellular functions. (2016)
  • Author(s): T. Suzuki, C. Huang and H. Fujihira
  • Journal Title: Gene Volume: 577 Issue: 1 Pages: 1-7
  • DOI: 10.1016/j.gene.2015.11.021
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Non-lysosomal degradation of phophorylated oligosaccharides is initiated by the cytosolic endo-beta-N-acetylglucosaminidase. (2016)
  • Author(s): Y. Harada, C Huang, S. Yamaki, N. Dohmae, and T. Suzuki
  • Journal Title: J. Biol. Chem. Volume: 291 Issue: 15 Pages: 8048-8058
  • DOI: 10.1074/jbc.m115.685313
  • Peer Reviewed
• [Journal Article] The cytoplasmic peptide:N-glycanase (Ngly1) - basic science encounters a human genetic disorder. (2015)
  • Author(s): Tadashi Suzuki
  • Journal Title: Journal of Biochemistry Volume: 157 Issue: 1 Pages: 23-34
  • DOI: 10.1093/jb/mvu068
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Endo-β-N-acetylglucosaminidase forms N-GlcNAc protein aggregates during ER-associated degradation in Ngly1-defective cells. (2015)
  • Author(s): Chengcheng Huang, Yoichiro Harada, Akira Hosom, Yuki Masahara-Negishi, Junichi Seino, Haruhiko Fujihira, Yoko Funakoshi, Takehiro Suzuki, Naoshi Dohmae, Tadashi Suzuki
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 112 Issue: 5 Pages: 1398-1403
  • DOI: 10.1073/pnas.1414593112
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Cytosolic-free oligosaccharides are predominantly generated by the degradation of dolichol-linked oligosaccharides in mammalian cells. (2015)
  • Author(s): Y. Harada, Y. Masahara-Negishi, and T. Suzuki.
  • Journal Title: Glycobiology Volume: 25 Issue: 11 Pages: 1196-1205
  • DOI: 10.1093/glycob/cwv055
  • Peer Reviewed
• [Journal Article] Forcible destruction of severely misfolded mammalian glycoproteins by the non-glycoprotein ERAD pathway. (2015)
  • Author(s): Ninagawa S, Okada T, Sumitomo Y, Horimoto S, Sugimoto T, Ishikawa T, Takeda S, Yamamoto T, Suzuki T, Kamiya Y, Kato K, Mori K.
  • Journal Title: The Journal of Cellular Biology Volume: 211 Issue: 4 Pages: 775-784
  • DOI: 10.1083/jcb.201504109
  • NAID: 120005672576
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Physiological and molecular functions of the cytosolic peptide:N-glycanase. (2015)
  • Author(s): Hiroto Hirayama, Akira Hosomi, and Tadashi Suzuki
  • Journal Title: Semin Cell Dev Biol. Volume: NA Pages: 110-120
  • DOI: 10.1016/j.semcdb.2014.11.009
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Generation and degradation of free asparagine-linked glycans. (2015)
  • Author(s): Yoichiro Harada, Hiroto Hirayama, and Tadashi Suzuki
  • Journal Title: Cell Mol. Life Sci. Volume: NA Issue: 13 Pages: 2509-2533
  • DOI: 10.1007/s00018-015-1881-7
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] エンドグリコシダーゼはヒト遺伝疾患の治療のターゲットになり得るか？ (2016)
  • Author(s): 鈴木 匡
  • Organizer: 2016年度日本農芸化学会大会
  • Place of Presentation: 札幌コンベンションセンター（北海道札幌市）
  • Year and Date: 2016-03-29
  • Invited
• [Presentation] ENGase as a potential drug target for NGLY1-deficiency. (2016)
  • Author(s): Tadashi Suzuki
  • Organizer: 7th Annual Rare Disease Day Symposium
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-02-27
  • Int'l Joint Research / Invited
• [Presentation] Battling with NGLY1-deficiency: can pure science cure a human genetic disorder ? (2016)
  • Author(s): Tadashi Suzuki
  • Organizer: The 3rd International Symposium on Glyco-Neuroscience
  • Place of Presentation: 淡路夢舞台国際会議場（兵庫県淡路市）
  • Year and Date: 2016-01-15
  • Int'l Joint Research / Invited
• [Presentation] Towards understanding the physiological role of non-lysosomal glycan catabolism. (2015)
  • Author(s): Tadashi Suzuki
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] Story of the cytoplasmic peptide:N-glycanase (Ngly1); update (2015)
  • Author(s): Tadashi Suzuki
  • Organizer: RIKEN-Academia Sinica Joint Conference on Chemical Biology
  • Place of Presentation: Taipei, Taiwan
  • Year and Date: 2015-10-15
  • Int'l Joint Research / Invited
• [Presentation] Story of Ngly1: beauty of basic research. (2015)
  • Author(s): Tadashi Suzuki
  • Organizer: 4th Austria/Japan Seminar on Comparative and Developmental Glycobiology
  • Place of Presentation: Split, Croatia
  • Year and Date: 2015-09-15
  • Int'l Joint Research / Invited
• [Presentation] 細胞質ペプチド：N-グリカナーゼ（Ngly1）の生理機能 (2015)
  • Author(s): 鈴木 匡
  • Organizer: 第34回日本糖質学会年会
  • Place of Presentation: 東京大学安田講堂（東京都文京区）
  • Year and Date: 2015-08-01
  • Invited
• [Presentation] 哺乳動物の脱糖鎖酵素（PNGase/ENGase）の生物学的重要性 (2015)
  • Author(s): 鈴木 匡
  • Organizer: 第1回 ENGase研究会
  • Place of Presentation: 理化学研究所 大河内ホール（埼玉県和光市）
  • Year and Date: 2015-07-31
  • Invited
• [Presentation] グライコフォームイメージングと糖タンパク質の細胞内動態解析 (2015)
  • Author(s): 鈴木 匡
  • Organizer: 第2回蛋白質工学研究会ワークショップ
  • Place of Presentation: あわぎんホール（徳島県徳島市）
  • Year and Date: 2015-06-23
  • Invited
• [Presentation] Story of Ngly1: an update. (2015)
  • Author(s): Tadashi Suzuki
  • Organizer: RIKEN-Max Planck Joint Research Center for Systems Chemical Biology The fourth Symposium
  • Place of Presentation: ニチイ学館 神戸ポートアイランドセンター（兵庫県神戸市）
  • Year and Date: 2015-05-12
  • Int'l Joint Research / Invited
• [Presentation] Story of the cytoplasmic peptide:N-glycanase (PNGase; Ngly1); basic science encounters human genetic disorder. (2015)
  • Author(s): Tadashi Suzuki
  • Organizer: GlycoCom
  • Place of Presentation: Banff, Canada
  • Year and Date: 2015-05-05
  • Int'l Joint Research / Invited
• [Presentation] Physiological functions of deglycosylating enzymes in mice. (2014)
  • Author(s): Haruhiko Fujihira, Yuki Masahara-Negishi, Masaru Tamura, Shigeharu Wakana, Chengcheng Huang, Gen Kondoh, Tadashi Yamashita, Toko Funakoshi, and Tadashi Suzuki
  • Organizer: Joint Meeting of the Society for Glycobiology and the Japanese Society of Carbohydrate Research
  • Place of Presentation: Honolulu (Hawaii, USA)
  • Year and Date: 2014-11-19
• [Presentation] The cytoplasmic PNGase (NGLY1): a 20-plus year scientific journey encounters a human genetic disorder. (2014)
  • Author(s): Tadashi Suzuki
  • Organizer: Joint Meeting of the Society for Glycobiology and the Japanese Society of Carbohydrate Research
  • Place of Presentation: Honolulu (Hawaii, USA)
  • Year and Date: 2014-11-18
  • Invited
• [Presentation] Functional studies of cytosolic deglycosylating enzyms in mammalian cells. (2014)
  • Author(s): Chengcheng Huang, Yoichiro Harada, Akira Hosomi, Yuki Masahara-Negishi, Junichi Seino, Haruhiko Fujihira, Yoko Funakoshi, Takehiro Suzuki, Naoshi Dohmae, and Tadashi Suzuki
  • Organizer: Joint Meeting of the Society for Glycobiology and the Japanese Society of Carbohydrate Research
  • Place of Presentation: Honolulu (Hawaii, USA)
  • Year and Date: 2014-11-18
• [Presentation] Physiological functions of deglycosylating enzymes in mice. (2014)
  • Author(s): Haruhiko Fujihira, Yuki Masahara-Negishi, Masaru Tamura, Shigeharu Wakana, Chengcheng Huang, Gen Kondoh, Tadashi Yamashita, Toko Funakoshi, and Tadashi Suzuki
  • Organizer: 2014 SFG & JSCR Joint Meeting Satellite Symposium II Glycans in Neuroscience
  • Place of Presentation: Honolulu (Hawaii, USA)
  • Year and Date: 2014-11-16
• [Presentation] Functional studies of cytosolic deglycosylating enzyms in mammalian cells. Physiological functions of deglycosylating enzymes in mice. (2014)
  • Author(s): Chengcheng Huang, Yoichiro Harada, Akira Hosomi, Yuki Masahara-Negishi, Junichi Seino, Haruhiko Fujihira, Yoko Funakoshi, Takehiro Suzuki, Naoshi Dohmae, and Tadashi Suzuki
  • Organizer: 2014 SFG & JSCR Joint Meeting Satellite Symposium II Glycans in Neuroscience
  • Place of Presentation: Honolulu (Hawaii, USA)
  • Year and Date: 2014-11-16
• [Presentation] マウスにおける糖鎖脱離酵素群の生理機能 (2014)
  • Author(s): 藤平 陽彦、根岸-正原 由紀、田村 勝、若菜 茂晴、黄 澄澄、近藤 玄、山下 匡、船越 陽子、鈴木 匡
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-16
• [Presentation] Functional analysis of cytosolic deglycosylating enzymes in mammalian cells. (2014)
  • Author(s): Chengcheng Huang, Yoichiro Harada, Akira Hosomi, Yuki Masahara-Negishi, Junichi Seino, Haruhiko Fujihira, Yoko Funakoshi, Takehiro Suzuki, Naoshi Dohmae, Tadashi Suzuki
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-16
• [Presentation] Story of the cytoplasmic peptide:N-glycanase -basic science meets a human genetic discorder. (2014)
  • Author(s): Tadashi Suzuki
  • Organizer: 25th Joint Glycobiology Meeting
  • Place of Presentation: Ghent (Belgium)
  • Year and Date: 2014-09-16
  • Invited
• [Presentation] 哺乳動物細胞の細胞質に局在する糖鎖脱離酵素の研究 (2014)
  • Author(s): 黄 澄澄、原田 陽一郎、細見 昭、正原（根岸）由紀、清野淳一、藤平陽彦、船越陽子、鈴木健裕、堂前直、鈴木 匡
  • Organizer: 第33回日本糖質学会年会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-08-12
• [Book] Sugar Chains (2015)
  • Author(s): Tadashi Suzuki, Kazuaki Ohtsubo, Naoyuki Taniguchi (eds)
  • Total Pages: 288
  • Publisher: Springer
• [Book] N-glycans and quality control of proteins. In Sugar Chains (T. Suzuki, K. Ohtsubo and N. Taniguchi eds.) (2015)
  • Author(s): Nobuko Hosokawa and Tadashi Suzuki
  • Publisher: Springer
• [Book] Overview of the quality control system for glycoproteins in the endoplasmic reticulum In Glycoscience: Biology and Medicine. (N. Taniguchi, T. Endo, G. W. Hart, P. Seeberger, C-H. Wong eds.). (2014)
  • Author(s): Tadashi Suzuki
  • Publisher: Springer