Study of recognition mechanism on glycan-biosynthesis in the Golgi by means of chemically synthesized homogeneous glycoproteins

• Project/Area Number: 17H01214
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bio-related chemistry
• Research Institution: Osaka University
• Principal Investigator: Kajihara Yasuhiro 大阪大学, 理学研究科, 教授 (50275020)
• Co-Investigator(Kenkyū-buntansha): 佐藤 あやの 岡山大学, ヘルスシステム統合科学研究科, 准教授 (40303002)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥39,650,000 (Direct Cost: ¥30,500,000、Indirect Cost: ¥9,150,000); Fiscal Year 2019: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000); Fiscal Year 2018: ¥12,480,000 (Direct Cost: ¥9,600,000、Indirect Cost: ¥2,880,000); Fiscal Year 2017: ¥14,690,000 (Direct Cost: ¥11,300,000、Indirect Cost: ¥3,390,000)
• Keywords: 糖タンパク質 / 複合型糖鎖 / 糖タンパク質生合成 / 分枝糖鎖 / ゴルジ体 / 糖鎖 / 蛋白質 / 糖 / ゴルジ装置

Outline of Final Research Achievements

A New chemical peptide-peptide coupling method was established and we used it for the synthesis of glycoproteins. Hydrogen-Deuterium exchange mass spectroscopy revealed that protein surface around N-glycans showed different deuterium pattern than that of other non-glycosylated protein surfaces. We demonstrated the insertion of folded native glycoprotein into the Golgi apparatus of the live cells. We found that biosynthesis of the antennary form of N-glycan seems to recognize a hydrophobic and hydrophilic pattern of protein surface by the use of mutation protocol of cytokine.

Academic Significance and Societal Importance of the Research Achievements

DNA、タンパク質についで、糖鎖は生体の第３の鎖として、その機能解明の研究が展開されてきた。
しかし、糖鎖は、天文学的な数の異性体を生じるため、その機能解明は困難を極めている。本研究では、タンパク質上の糖鎖が、周りの水と相互作用をすることで、タンパク質の機能を高めているという現象を見出すことができた。その結果、糖鎖と水の相互作用に着目することでこれまで理解できていなかった糖鎖をもつタンパク質の機能、および糖鎖がタンパク質へ与える影響を研究できる基盤ができた。

Research Products

• [Journal Article] Total Chemical Synthesis of a Nonfibrillating Human Glycoinsulin (2020)
  • Author(s): M. A. Hossain, R. Okamoto, J. A. Karas, P. Praveen, M. Liu, B. E. Forbes, J. D. Wade, Y. Kajihara
  • Journal Title: J. Am. Chem. Soc. Volume: 142 Issue: 3 Pages: 1164-1169
  • DOI: 10.1021/jacs.9b11424
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Chemical Modification of the N Termini of Unprotected Peptides for Semisynthesis of Modified Proteins by Utilizing a Hydrophilic Protecting Group (2019)
  • Author(s): C. Chandrashekar, R.Okamoto, M. Izumi,Y. Kajihara
  • Journal Title: Chem. Eur. J. Volume: 25 Issue: 43 Pages: 10197-10203
  • DOI: 10.1002/chem.201901778
  • Peer Reviewed
• [Journal Article] The Golgin Protein Giantin Regulates Interconnections Between Golgi Stacks (2019)
  • Author(s): Satoh Ayano、Hayashi-Nishino Mitsuko、Shakuno Takuto、Masuda Junko、Koreishi Mayuko、Murakami Runa、Nakamura Yoshimasa、Nakamura Toshiyuki、Abe-Kanoh Naomi、Honjo Yasuko、Malsam Joerg、Yu Sidney、Nishino Kunihiko
  • Journal Title: Frontiers in Cell and Developmental Biology Volume: 7 Pages: 160-160
  • DOI: 10.3389/fcell.2019.00160
  • NAID: 120006733903
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regioselective α-Peptide Bond Formation Through the Oxidation of Amino Thioacids (2019)
  • Author(s): Okamoto Ryo、Haraguchi Takuya、Nomura Kota、Maki Yuta、Izumi Masayuki、Kajihara Yasuhiro
  • Journal Title: Biochemistry Volume: 58 Issue: 12 Pages: 1672-1678
  • DOI: 10.1021/acs.biochem.8b01239
  • Peer Reviewed
• [Journal Article] Semisynthesis of Complex-Type Biantennary Oligosaccharides Containing Lactosamine Repeating Units from a Biantennary Oligosaccharide Isolated from a Natural Source (2018)
  • Author(s): Yuta Maki, Takanori Mima, Ryo Okamoto, Masayuki Izumi, and Yasuhiro Kajihara
  • Journal Title: J. Org. Chem. Volume: 83 Issue: 1 Pages: 443-451
  • DOI: 10.1021/acs.joc.7b02485
  • Peer Reviewed
• [Journal Article] Chemical synthesis of glycoproteins with the specific installation of gradient enriched 15N-labeled amino acids for getting insight into glycoprotein behavior (2017)
  • Author(s): Nguyen Minh Hien, Masayuki Izumi, Hajime Sato, Ryo Okamoto, Yasuhiro Kajihara
  • Journal Title: Chemistry A European Journal Volume: 23 Issue: 27 Pages: 6579-6585
  • DOI: 10.1002/chem.201606049
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Presentation] ゴルジ体における糖鎖分枝修飾の機能解明に向けたdi-GlcNAc糖鎖を有するCholera Toxin B-subunit (2020)
  • Author(s): 梶原 康宏, 岡本 亮, 真木 勇太, 佐藤 あやの, 吉井 優也, 川田 一稀
  • Organizer: 本化学会 第100春季年会
• [Presentation] 光感受性官能基を持つ新規N結合型糖ペプチドの合成研究 (2019)
  • Author(s): 石原 薫、岡 彩惠、 真木勇太 、 岡本 亮、 梶原康宏
  • Organizer: 第38回日本糖質学会年会
• [Presentation] 非天然型糖鎖を有する不凍糖タンパク質誘導体の化学合成とその機能評価 (2019)
  • Author(s): 芝田 大之・真木 勇太・深見 大地・津田 栄・梶原 康宏・岡本 亮
  • Organizer: 第20回関西グライコサイエンスフォーラム
• [Presentation] Elucidation of sugar functions of antifreeze glycoprotein (2019)
  • Author(s): SHIBATA, Hiroyuki; ORII, Ryo; MAKI, Yuta; FUKAMI, Daichi; TSUDA, Sakae; KAJIHARA,
  • Organizer: 日本化学会第100春季年会
• [Presentation] Functions of N-Glycan on Protein (2019)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 8 th Chemical Protein Synthesis
  • Int'l Joint Research / Invited
• [Presentation] Functions of N-Glycan on Protein (2019)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 2019 American chemical Society national meeting
  • Int'l Joint Research / Invited
• [Presentation] Chemical Synthesis of Homogeneous Glycoprotein Having a Tri-antennary Sialyloligosaccharide. (2019)
  • Author(s): Yasuhiro Kajihara, Yuta Maki
  • Organizer: 7Th MODERN SOLID PHASE PEPTIDE SYNTHESIS & ITS APPLICATIONS SYMPOSIUM
  • Int'l Joint Research / Invited
• [Presentation] Semi-synthesis of Homogeneous Glycoproteins, 13 th Australian Peptide Conference (2019)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 13 th Australian Peptide Conference
  • Int'l Joint Research / Invited
• [Presentation] Functions of N-Glycan on Protein (2019)
  • Author(s): Yasuhiro Kajihara
  • Organizer: Emil Fischer memoriam symposium
  • Int'l Joint Research / Invited
• [Presentation] Functions of N-Glycan on Protein (2019)
  • Author(s): Yasuhiro Kajihara
  • Organizer: IUPAC international symposium Bioorganic chemistry
  • Int'l Joint Research / Invited
• [Presentation] 炭水化物研究 (2019)
  • Author(s): 梶原康宏
  • Organizer: 38回日本糖質学会年会
  • Invited
• [Presentation] 糖タンパク質糖鎖の普遍的機能 (2019)
  • Author(s): 梶原康宏
  • Organizer: 17回糖鎖科学コンソーシアムシンポジウム
  • Invited
• [Presentation] ゴルジ体ゾーンの形成にゴルジンタンパク質であるGiantinは関与するか (2019)
  • Author(s): 佐藤あやの、杓野拓斗、西野林美都子、西野邦彦
  • Organizer: 第19回日本蛋白質科学会年会
• [Presentation] コレラトキシンBサブユニットを利用した機能性ペプチドの小胞体への特異的送達法の開発 (2019)
  • Author(s): 佐藤あやの、吉井優也、寺尾-是石真友子
  • Organizer: 第38回日本糖質学会年会
• [Presentation] コレラトキシンBサブユニットを利用した機能性ペプチドの細胞小器官への送達 (2019)
  • Author(s): 吉井優也、寺尾-是石真友子、佐藤あやの
  • Organizer: 第71回日本生物工学会大会
• [Presentation] Semisynthesis of Complex-Type Biantennary Oligosaccharides Containing Lactosamine Repeating Units from a Biantennary Oligosaccharide Isolated from a Natural Source (2018)
  • Author(s): Yuta Maki, Ryo Okamoto, Masayuki Izumi, Yasuhiro Kajihara
  • Organizer: 29th International Carbohydrate Symposium
  • Int'l Joint Research
• [Presentation] Chemical Synthesis of Homogeneous Sialylglycoproteins (2018)
  • Author(s): Yasuhiro Kajihara
  • Organizer: Sialoglyco2018
  • Int'l Joint Research / Invited
• [Presentation] N-GLYCANS ON PROTEINS (2018)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 29th International Carbohydrate Symposium
  • Int'l Joint Research / Invited
• [Presentation] 糖タンパク質の精密化学合成を利用する糖鎖機能解明 (2018)
  • Author(s): 梶原康宏
  • Organizer: 日本農芸化学会 中部支部 第183例会
  • Invited
• [Presentation] 糖タンパク質の精密化学合成 (2018)
  • Author(s): 梶原康宏
  • Organizer: 日本生物学会2018
  • Invited
• [Presentation] ゴルジ体由来のN-アセチルガラクトサミン転移酵素の基質特異性 (2018)
  • Author(s): 岩木 ゆか、真木 勇太、岡本 亮、梶原 康宏
  • Organizer: 日本化学会第98春季年会
• [Presentation] Synthesis of Glycoconjugates for Elucidation of Oligosaccharide Functions (2017)
  • Author(s): Yasuhiro Kajihara
  • Organizer: Gordon Research Conference (Carbohydrate)
  • Int'l Joint Research / Invited
• [Presentation] New Approach for Chemical Synthesis of Glycoproteins (2017)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 7th Chemical Protein Synthesis Meeting
  • Int'l Joint Research / Invited
• [Presentation] Synthesis of sialyltransferase (2017)
  • Author(s): Yasuhiro Kajihara
  • Organizer: 9th Asian Community of Glycoscience and Glycotechnology Conference
  • Int'l Joint Research
• [Presentation] ゴルジ体における糖鎖構造制御システムの解明研究 (2017)
  • Author(s): 岡 彩惠 、萱原 沙耶、真木 勇太、岡本 亮、和泉 雅之 、梶原 康宏
  • Organizer: 第36回 日本糖質学会年会
• [Presentation] 第36回 日本糖質学会年会 (2017)
  • Author(s): 西川 理恵 、真木 勇太、岡本 亮、和泉 雅之 、梶原 康宏
  • Organizer: 第36回 日本糖質学会年会
• [Book] Advances in Experimental Medicine and Biology (2019)
  • Author(s): Amazaki, Yoko; Nguyen, Hien Minh; Okamoto, Ryo; Maki, Yuta; Kajihara, Yasuhiro
  • Total Pages: 19
  • Publisher: Springer
• [Patent(Industrial Property Rights)] ペプチドチオエステル、ペプチドの新規製造方法 (2018)
  • Inventor(s): 梶原康宏、岡本亮、真木勇太, 西川理恵、村瀬健文、他２名
  • Industrial Property Rights Holder: 糖鎖工学研究所
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-226094
  • Filing Date: 2018