Structural studies on the variety and universality of GlcNAc transferases

• Project/Area Number: 20K06575
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Osaka University
• Principal Investigator: Nagae Masamichi 大阪大学, 微生物病研究所, 助教 (60619873)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 構造生物学 / 糖鎖生物学

Outline of Research at the Start

本研究のコンセプトは、ごくありふれた糖鎖であるNアセチルグルコサミン（GlcNAc）が引き起こす多様な生理応答をGlcNAcを転移する酵素の立体構造を通して明らかにすることである。GlcNAcはグルコースから合成され、細胞内外に豊富に存在する糖質である。GlcNAcは様々なタンパク質に様々な様式で付加され、多様な機能を発揮する。これを担うGlcNAc転移酵素は進化的にかけ離れたものがいくつも存在する。申請者はこの点に着目して、生物がどのような過程を経てGlcNAcを有用に利用できるようになったのかを明らかにしたい。

Outline of Final Research Achievements

N-acetylglucosamine (GlcNAc) is synthesized by glucose and one of the most abundant monosaccharide in our body. GlcNAc is attached to glycoproteins via various linkages by multiple GlcNAc transferases. However, few studies focused on the commonality and uniqueness of these GlcNAc addition from the structural points of view. In this study, we performed structural studies on three GlcNAc transferases, GnT-IVa, GnT-V, and EOGT, to elucidate their substrate recognition and reaction mechanisms. As a result, we found a new functional domain in GnT-IVa. We also identified residues involved in the substrate glycoprotein of GnT-V. Furthermore, we succeeded in crystallizing EOGT.

Academic Significance and Societal Importance of the Research Achievements

GlcNAc転移酵素は多様なファミリーから構成される糖転移酵素群であり、その生理機能は多岐に渡っており、関連する疾患も複数存在する。本研究が対象としたGnT-IVaは膵臓のβ細胞上のグルコースの取り込みを調節することが知られており、II型糖尿病への関与が報告されている。またGnT-Vの過剰発現は細胞のがん化に関与することが知られている。さらにEOGTは難病であるAdams-Oliver症候群の原因遺伝子である。本研究はこれらの酵素の共通性と独自性を明らかにすることを目指しており、その成果はこれらの酵素の制御、ひいては疾患の制御につながることが期待される。

Research Products

• [Journal Article] Discovery of a lectin domain that regulates enzyme activity in mouse N-acetylglucosaminyltransferase-IVa (MGAT4A) (2022)
  • Author(s): Nagae Masamichi、Hirata Tetsuya、Tateno Hiroaki、Mishra Sushil K.、Manabe Noriyoshi、Osada Naoko、Tokoro Yuko、Yamaguchi Yoshiki、Doerksen Robert J.、Shimizu Toshiyuki、Kizuka Yasuhiko
  • Journal Title: Communications Biology Volume: 5 Issue: 1 Pages: 695-695
  • DOI: 10.1038/s42003-022-03661-w
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Examination of differential glycoprotein preferences of N-acetylglucosaminyltransferase-IV isozymes a and b (2022)
  • Author(s): Osada Naoko、Nagae Masamichi、Nakano Miyako、Hirata Tetsuya、Kizuka Yasuhiko
  • Journal Title: Journal of Biological Chemistry Volume: 298 Issue: 9 Pages: 102400-102400
  • DOI: 10.1016/j.jbc.2022.102400
  • Peer Reviewed / Open Access
• [Journal Article] The stem region of α1,6-fucosyltransferase FUT8 is required for multimer formation but not catalytic activity (2022)
  • Author(s): Tomida Seita、Nagae Masamichi、Kizuka Yasuhiko
  • Journal Title: Journal of Biological Chemistry Volume: 298 Issue: 12 Pages: 102676-102676
  • DOI: 10.1016/j.jbc.2022.102676
  • Peer Reviewed / Open Access
• [Journal Article] Symbiotic bacteria-dependent expansion of MR1-reactive T cells causes autoimmunity in the absence of Bcl11b (2022)
  • Author(s): Shibata Kensuke、Motozono Chihiro、Nagae Masamichi、Shimizu Takashi、Ishikawa Eri、Motooka Daisuke、Okuzaki Daisuke、Izumi Yoshihiro、Takahashi Masatomo、Fujimori Nao、Wing James B.、Hayano Takahide、Asai Yoshiyuki、Bamba Takeshi、Ogawa Yoshihiro、Furutani-Seiki Makoto、Shirai Mutsunori、Yamasaki Sho
  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 6948-6948
  • DOI: 10.1038/s41467-022-34802-8
  • Peer Reviewed / Open Access
• [Journal Article] Self-referential immune recognition through C-type lectin receptors (2022)
  • Author(s): Guenther Carla、Nagae Masamichi、Yamasaki Sho
  • Journal Title: Adv. Immunol. Volume: 156 Pages: 1-23
  • DOI: 10.1016/bs.ai.2022.09.001
  • ISBN: 9780323989473
  • Peer Reviewed
• [Journal Article] Human Dectin-1 is O-glycosylated and serves as a ligand for C-type lectin receptor CLEC-2 (2022)
  • Author(s): Haji Shojiro、Ito Taiki、Guenther Carla、Nakano Miyako、Shimizu Takashi、Mori Daiki、Chiba Yasunori、Tanaka Masato、Mishra Sushil K、Willment Janet A、Brown Gordon D、Nagae Masamichi、Yamasaki Sho
  • Journal Title: eLife Volume: 11
  • DOI: 10.7554/elife.83037
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] N-acetylglucosaminyltransferase-V requires a specific noncatalytic luminal domain for its activity toward glycoprotein substrates. (2022)
  • Author(s): Osuka RF, Hirata T, Nagae M, Nakano M, Shibata H, Okamoto R, Kizuka Y.
  • Journal Title: J. Biol. Chem. Volume: 298 Issue: 3 Pages: 101666-101666
  • DOI: 10.1016/j.jbc.2022.101666
  • Peer Reviewed / Open Access
• [Journal Article] Structure-based design of UDP-GlcNAc analogs as candidate GnT-V inhibitors (2022)
  • Author(s): Vibhute Amol M.、Tanaka Hide-nori、Mishra Sushil K.、Osuka Reina F.、Nagae Masamichi、Yonekawa Chizuko、Korekane Hiroaki、Doerksen Robert J.、Ando Hiromune、Kizuka Yasuhiko
  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1866 Issue: 6 Pages: 130118-130118
  • DOI: 10.1016/j.bbagen.2022.130118
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Identification of conserved SARS-CoV-2 spike epitopes that expand public cTfh clonotypes in mild COVID-19 patients (2021)
  • Author(s): Lu X, Hosono Y, Nagae M, Ishizuka S, Ishikawa E, Motooka D, Ozaki Y, Sax N, Maeda Y, Kato Y, Morita T, Shinnakasu R, Inoue T, Onodera T, Matsumura T, Shinkai M, Sato T, Nakamura S, Mori S, Kanda T, Nakayama EE, Shioda T, Kurosaki T, Takeda K, Kumanogoh A, Arase H, Nakagami H, Yamashita K, Takahashi Y, Yamasaki S.
  • Journal Title: J Exp Med. Volume: 218 Issue: 12
  • DOI: 10.1084/jem.20211327
  • Peer Reviewed / Open Access
• [Journal Article] 3D Structure and Function of Glycosyltransferases Involved in N-glycan Maturation (2020)
  • Author(s): Nagae Masamichi、Yamaguchi Yoshiki、Taniguchi Naoyuki、Kizuka Yasuhiko
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 2 Pages: 437-437
  • DOI: 10.3390/ijms21020437
  • Peer Reviewed / Open Access
• [Journal Article] Structural insight into the recognition of pathogen-derived phosphoglycolipids by C-type lectin receptor DCAR (2020)
  • Author(s): Omahdi Zakaria、Horikawa Yuto、Nagae Masamichi、Toyonaga Kenji、Imamura Akihiro、Takato Koichi、Teramoto Takamasa、Ishida Hideharu、Kakuta Yoshimitsu、Yamasaki Sho
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 17 Pages: 5807-5817
  • DOI: 10.1074/jbc.ra120.012491
  • Peer Reviewed
• [Journal Article] The SH3 domain in the fucosyltransferase FUT8 controls FUT8 activity and localization and is essential for core fucosylation (2020)
  • Author(s): Tomida Seita、Takata Misaki、Hirata Tetsuya、Nagae Masamichi、Nakano Miyako、Kizuka Yasuhiko
  • Journal Title: Journal of Biological Chemistry Volume: 295 Issue: 23 Pages: 7992-8004
  • DOI: 10.1074/jbc.ra120.013079
  • Peer Reviewed / Open Access
• [Journal Article] Differential Labeling of Glycoproteins with Alkynyl Fucose Analogs (2020)
  • Author(s): Ma Chenyu、Takeuchi Hideyuki、Hao Huilin、Yonekawa Chizuko、Nakajima Kazuki、Nagae Masamichi、Okajima Tetsuya、Haltiwanger Robert S.、Kizuka Yasuhiko
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 17 Pages: 6007-6007
  • DOI: 10.3390/ijms21176007
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Recognition of glycan and protein substrates by N-acetylglucosaminyltransferase-V (2020)
  • Author(s): Hirata Tetsuya、Nagae Masamichi、Osuka Reina F.、Mishra Sushil K.、Yamada Mayumi、Kizuka Yasuhiko
  • Journal Title: Biochimica et Biophysica Acta (BBA) - General Subjects Volume: 1864 Issue: 12 Pages: 129726-129726
  • DOI: 10.1016/j.bbagen.2020.129726
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] True significance of N-acetylglucosaminyltransferases GnT-III, V and α1,6 fucosyltransferase in epithelial-mesenchymal transition and cancer (2020)
  • Author(s): Taniguchi Naoyuki、Ohkawa Yuki、Maeda Kento、Harada Yoichiro、Nagae Masamichi、Kizuka Yasuhiko、Ihara Hideyuki、Ikeda Yoshitaka
  • Journal Title: Molecular Aspects of Medicine Volume: in press Pages: 100905-100905
  • DOI: 10.1016/j.mam.2020.100905
  • Peer Reviewed
• [Journal Article] Structural studies of reelin N-terminal region provides insights into a unique structural arrangement and functional multimerization (2020)
  • Author(s): Nagae Masamichi、Suzuki Kei、Yasui Norihisa、Nogi Terukazu、Kohno Takao、Hattori Mitsuharu、Takagi Junichi
  • Journal Title: The Journal of Biochemistry Volume: mvaa144 Issue: 5 Pages: 1-41
  • DOI: 10.1093/jb/mvaa144
  • NAID: 40022623693
  • Peer Reviewed
• [Presentation] レクチン受容体による結晶性物質の認識機構に関する構造基盤の解明 (2022)
  • Author(s): 長江雅倫、森正太郎、山﨑晶
  • Organizer: 第41回日本糖質学会年会
• [Presentation] Structural basis of antigen recognition by distinct public TCRs recognizing an identical epitope conserved across SARS-CoV-2 variants (2022)
  • Author(s): Masamichi Nagae, Shotaro Mori, Taiki Ito, Xiuyuan, Lu, Sho Yamasaki
  • Organizer: 第51回日本免疫学会年会