Synthetic study of the O-glycoprotein using the solubilizing method

• Project/Area Number: 15K21124
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Bio-related chemistry; Biomolecular chemistry
• Research Institution: Osaka University
• Principal Investigator: Asahina Yuya 大阪大学, たんぱく質研究所, 助教 (10737232)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 糖ペプチド / 糖タンパク質 / 極性反転 / 可溶化 / ペプチドライゲーション / 4-ピコリル基（Pic） / インターロイキン－２ / シアル酸 / チオエステル法 / 親水性保護基 / 極性反転保護 / O-結合型糖鎖 / N-アルキルシステイン

Outline of Final Research Achievements

In this study, we successfully established a novel solubilizing method using 4-picolyl (Pic) group, which realized the synthesis of the glycoprotein, human interleukin-2 (IL-2), containing C-terminal poor solubility regions. Pic ester was introduced to the carboxy group of Glu residues of the C-terminal region, effectively solubilizing the poor solubility peptide. In the next step, the peptide ligation using the one-pot thioester method was performed to obtain the entire polypeptide. After the polypeptide was lead to the final product via deprotection and folding, the synthetic one was investigated by CD analysis and T-cell proliferation assay, which indicated the success of the chemical synthesis of IL-2 having homogeneous sugar chain.

Research Products

• [Journal Article] One-pot native chemical ligation by combination of two orthogonal thioester precursors (2017)
  • Author(s): Y. Asahina, Toru Kawakami and Hironobu Hojo
  • Journal Title: Chem. Commun Volume: 53 Issue: 13 Pages: 2114-2117
  • DOI: 10.1039/c6cc10243c
  • Peer Reviewed
• [Journal Article] Development of the One-pot Ligation by Using Two Orthogonal Thioesterification Devices (2017)
  • Author(s): Yuya Asahina, Toru Kawakami, Hironobu Hojo
  • Journal Title: Peptide Science (Proceeding) Volume: -
  • Peer Reviewed
• [Journal Article] Synthesis of Interleukin-2 Having Core 1 Type Disaccharide by the Reverse Polarity Protection Strategy (2016)
  • Author(s): Rina Fujimoto, Yuya Asahina, Shinobu Komiya, Ami Ohagi, Hiroko Tamagaki, Katsuhiro Nakagawa, Takashi Sato, Shizuo Akira, Toshifumi Takao, Akira Ishii, Yohsiaki Nakahara, Hironobu, Hojo
  • Journal Title: Peptide Science 2015: H. Hojo, T. Inazu, H. Katayama (Eds.) Volume: - Pages: 119-122
  • Peer Reviewed
• [Journal Article] Synthesis of O-Linked Glycopeptide Thioester by the 4-Methylbenzyl Protection Strategy (2016)
  • Author(s): Naoki Takeda, Yuya Asahina, Rina Fujimoto, Akemi Suzuki, Hironobu Hojo
  • Journal Title: Peptide Science 2015: H. Hojo, T. Inazu, H. Katayama (Eds.) Volume: - Pages: 123-124
  • Peer Reviewed
• [Journal Article] Study on a Preparation of Prolyl Thioester by the N-Alkylcysteine Method (2016)
  • Author(s): Ami Ohagi, Yuya Asahina, Hironobu Hojo
  • Journal Title: Peptide Science 2015: H. Hojo, T. Inazu, H. Katayama (Eds.) Volume: - Pages: 135-136
  • Peer Reviewed
• [Journal Article] Chemical Synthesis of O-Glycosylated Human Interleukin-2 by the Reverse Polarity Protection Strategy (2015)
  • Author(s): Yuya Asahina, Shinobu Komiya, Ami Ohagi, Rina Fujimoto,Hiroko Tamagaki, Katsuhiro Nakagawa, Takashi Sato,Shizuo Akira, Toshifumi Takao,Akira Ishii, Yoshiaki Nakahara, Hironobu Hojo
  • Journal Title: Angewandte Chemie International Edition Volume: 54 Issue: 28 Pages: 8226-8230
  • DOI: 10.1002/anie.201501847
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Chemoenzymatic Synthesis of Glycoprotein Using the Peptide Ligation Chemistry (2015)
  • Author(s): Yuya Asahina
  • Journal Title: Trends in Glycoscience and Glycotechnology Volume: 27 Issue: 157 Pages: E15-E19
  • DOI: 10.4052/tigg.1414.4E
  • NAID: 130005099230
  • ISSN: 0915-7352, 1883-2113
  • Peer Reviewed
• [Journal Article] Chemoenzymatic Synthesis of Glycoprotein Using the Peptide Ligation Chemistry (Jpn. Ed.) (2015)
  • Author(s): 朝比奈 雄也
  • Journal Title: Trends in Glycoscience and Glycotechnology Volume: 27 Issue: 157 Pages: J15-J20
  • DOI: 10.4052/tigg.1414.4J
  • NAID: 130005099227
  • ISSN: 0915-7352, 1883-2113
  • Peer Reviewed
• [Presentation] 直交型チオエステル化素子を利用した ヒトヒストンH4の化学合成 (2017)
  • Author(s): 朝比奈雄也、川上徹、北條裕信
  • Organizer: 日本農芸化学会２０１７年会
  • Place of Presentation: 京都女子大学（京都府、京都市）
  • Year and Date: 2017-03-17
• [Presentation] カベオリン-1の化学合成研究及び細胞膜内ドメインの構造解析 (2017)
  • Author(s): 藤本梨菜、武居俊樹、朝比奈雄也、佐藤毅、北條裕信
  • Organizer: 日本農芸化学会２０１７年会
  • Place of Presentation: 京都女子大学（京都府、京都市）
  • Year and Date: 2017-03-17
• [Presentation] TFA感受性保護基を用いたシアリル糖ペプチドの合成 (2017)
  • Author(s): 竹田直樹、武居俊樹、朝比奈雄也、北條裕信
  • Organizer: 日本農芸化学会２０１７年会
  • Place of Presentation: 京都女子大学（京都府、京都市）
  • Year and Date: 2017-03-17
• [Presentation] STUDY FOR DEVELOPMENT OF THE ONE-POT LIGATION BY USING TWO ORTHOGONAL THIOESTERIFICATION DEVICES (2016)
  • Author(s): Yuya Asahina, Toru Kawakami, Hironobu Hojo
  • Organizer: 第５３回ペプチド討論会
  • Place of Presentation: 京都テルサ（京都府、京都市）
  • Year and Date: 2016-10-26
• [Presentation] Total Synthesis of Human Interleukin-2 Having Core 1 Disaccharide by the Reverse Polarity Protection Strategy (2016)
  • Author(s): Yuya Asahina, Shinobu Komiya, Ami Ohagi, Rina Fujimoto, Hiroko Tamagaki, Katsuhiro Nakagawa, Takashi Sato, Shizuo Akira, Toshifumi Takao, Akira Ishii, Yoshiaki Nakahara, Hironobu Hojo
  • Organizer: 8th International Peptide Symposium
  • Place of Presentation: ライプツィヒ大学（ライプツィヒ、ドイツ）
  • Year and Date: 2016-09-04
  • Int'l Joint Research
• [Presentation] 4-メチルベンジル基を用いたalpha-シアリル糖ペプチドの合成研究 (2016)
  • Author(s): 竹田直樹、武居俊樹、朝比奈雄也、北條裕信
  • Organizer: 第３５回日本糖質学会年会
  • Place of Presentation: 高知市文化プラザ かるぽーと（高知県高知市）
  • Year and Date: 2016-09-01
• [Presentation] STUDY ON A PREPARATION OF PROLYL THIOESTER BY THE N- ALKYLCYSTEINE METHOD (2015)
  • Author(s): Ami Ohagi, Yuya Asahina, Hironobu Hojo
  • Organizer: 第52回日本ペプチド討論会
  • Place of Presentation: 平塚公民館（ 神奈川県平塚市 ）
  • Year and Date: 2015-11-16
• [Presentation] TOTAL SYNTHESIS OF INTERLEUKIN-2 HAVING CORE 1 TYPE DISACCHARIDE BY THE REVERSE POLARITY PROTECTION STRATEGY (2015)
  • Author(s): Rina Fujimoto, Yuya Asahina, Shinobu Komiya, Ami Ohagi, Hiroko Tamagaki, Katsuhiro Nakagawa, Takashi Sato, Shizuo Akira, Toshifumi Takao, Akira Ishii, Yoshiaki Nakahara, Hironobu Hojo
  • Organizer: 第52回日本ペプチド討論会
  • Place of Presentation: 平塚公民館（ 神奈川県平塚市 ）
  • Year and Date: 2015-11-16
• [Presentation] SYNTHESIS OF O-LINKED GLYCOPEPTIDE THIOESTER BY THE 4- METHYLBENZYL PROTECTION STRATEGY (2015)
  • Author(s): Naoki Takeda, Yuya Asahina, Rina Fujimoto, Akemi Suzuki, Hironobu Hojo
  • Organizer: 第52回日本ペプチド討論会
  • Place of Presentation: 平塚公民館（ 神奈川県平塚市 ）
  • Year and Date: 2015-11-16
• [Presentation] O-結合型糖鎖を有するインターロイキン－２の化学合成 (2015)
  • Author(s): 朝比奈雄也、小宮忍、大萩亜美、藤本梨菜、玉垣裕子、中川勝博、佐藤荘、審良静男、石井彰、高尾敏文、中原義昭、北條裕信
  • 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: 2015-07-31
• [Remarks] 2種類のチオエステル化前駆体を利用したワンポットNCL法
  • URL: http://www.protein.osaka-u.ac.jp/organic/res_desc/index.html#cpenac
• [Remarks] IL2の化学合成
  • URL: http://www.protein.osaka-u.ac.jp/organic/res_desc/index.html#IL2