The development of analytical method for sialylated glycans and glycoproteins by aminolysis-SALSA

• Project/Area Number: 19K15597
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 34020:Analytical chemistry-related
• Research Institution: Hokkaido University
• Principal Investigator: Hanamatsu Hisatoshi 北海道大学, 医学研究院, 特任助教 (70734185)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 糖鎖解析 / シアル酸 / 質量分析 / グライコミクス / 糖鎖

Outline of Research at the Start

核酸、タンパク質に次ぐ第三の生命鎖である糖鎖は多くの疾患や生命現象に関与している。糖鎖を構成する糖の一つであるシアル酸は非常に多様な結合様式（α2,3、α2,6、α2,8）を有して存在しており、その結合様式の違いが免疫細胞の機能やウイルス感染に影響を与えることが知られている。本研究では、申請者らが開発した α2,3 シアル酸開環アミノリシスを基盤としたシアル酸結合様式特異的修飾法 (Aminolysis-SALSA) を応用することで、安定同位体試薬を用いた糖鎖比較解析法、シアル酸結合糖タンパク質の精製・濃縮法などの開発を目指す。

Outline of Final Research Achievements

In order to develop novel analytical methods for sialylated glycans and glycoproteins/peptides, we investigated the application of sialic acid linkage-specific alkylamidation via lactone ring-opening aminolysis, termed as aminolysis-SALSA. In this study, we developed comparative and quantitative glycomic analysis by isotope labeling of α2,3-linked sialic acid residues and purification and concentration method of sialylated glycoproteins/peptides using biotin-derivatized amine reagents.Moreover, we found that α2,3 sialyl linkage-specific amidation of esterified sialoglycans can be achieved via an intramolecular lactone, and reported as lactone-driven ester-to-amide derivatization method.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した、安定同位体試薬を用いた α2,3シアル酸特異的修飾法、ビオチン誘導体化アミン試薬を用いたシアリル化糖タンパク質/ペプチドの修飾法は、疾患糖鎖バイオマーカー探索、投薬による治療効果のモニタリング、血液、細胞、組織など多様な生体サンプルからの α2,3シアル酸を有する糖タンパク質/ペプチド特異的な精製・濃縮などへの応用が期待される。また、エステル-アミド交換法はシアル酸結合様式の識別だけではなく、分岐構造を持つ糖鎖の識別も可能であり、より詳細な糖鎖構造異性体の識別に繋がる。

Research Products

• [Journal Article] A defined glycosylation regulatory network modulates total glycome dynamics during pluripotency state transition (2021)
  • Author(s): Pecori Federico、Yokota Ikuko、Hanamatsu Hisatoshi、Miura Taichi、Ogura Chika、Ota Hayato、Furukawa Jun-ichi、Oki Shinya、Yamamoto Kazuo、Yoshie Osamu、Nishihara Shoko
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 1276-1276
  • DOI: 10.1038/s41598-020-79666-4
  • Peer Reviewed / Open Access
• [Journal Article] The technical developments of analytical methods for the primary structure of glycans (2020)
  • Author(s): 花松 久寿、古川 潤一
  • Journal Title: 生化学 Volume: 92 Issue: 3 Pages: 360-368
  • DOI: 10.14952/SEIKAGAKU.2020.920360
  • ISSN: 0037-1017
  • Year and Date: 2020-06-25
• [Journal Article] Lactone-Driven Ester-to-Amide Derivatization for Sialic Acid Linkage-Specific Alkylamidation (2020)
  • Author(s): Jun-Ichi Furukawa 1, Hisatoshi Hanamatsu 2, Takashi Nishikaze 3, Hiroshi Manya 4, Nobuaki Miura 5, Hirokazu Yagi 6, Ikuko Yokota 1, Keiko Akasaka-Manya 4, Tamao Endo 4, Motoi Kanagawa 7 8, Norimasa Iwasaki 1, Koichi Tanaka 3
  • Journal Title: Anal Chem . Volume: 3 Issue: 21 Pages: 14383-14392
  • DOI: 10.1021/acs.analchem.0c02209
  • Peer Reviewed / Open Access
• [Journal Article] Toolbox Accelerating Glycomics (TAG): Glycan Annotation from MALDI-TOF MS Spectra and Mapping Expression Variation to Biosynthetic Pathways (2020)
  • Author(s): Miura Nobuaki、Hanamatsu Hisatoshi、Yokota Ikuko、Okada Kazue、Furukawa Jun-Ichi、Shinohara Yasuro
  • Journal Title: Biomolecules Volume: 10 Issue: 10 Pages: 1383-1383
  • DOI: 10.3390/biom10101383
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Alterations of Glycosphingolipid Glycans and Chondrogenic Markers during Differentiation of Human Induced Pluripotent Stem Cells into Chondrocytes (2020)
  • Author(s): Xu Liang、Hanamatsu Hisatoshi、Homan Kentaro、Onodera Tomohiro、Miyazaki Takuji、Furukawa Jun-ichi、Hontani Kazutoshi、Tian Yuan、Baba Rikiya、Iwasaki Norimasa
  • Journal Title: Biomolecules Volume: 10 Issue: 12 Pages: 1622-1622
  • DOI: 10.3390/biom10121622
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mucin-type O-glycosylation controls pluripotency in mouse embryonic stem cells via Wnt receptor endocytosis (2020)
  • Author(s): Pecori Federico、Akimoto Yoshihiro、Hanamatsu Hisatoshi、Furukawa Jun-ichi、Shinohara Yasuro、Ikehara Yuzuru、Nishihara Shoko
  • Journal Title: Journal of Cell Science Volume: 133 Issue: 20 Pages: 245845-245845
  • DOI: 10.1242/jcs.245845
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comparative Glycomic Analysis of Sialyl Linkage Isomers by Sialic Acid Linkage-Specific Alkylamidation in Combination with Stable Isotope Labeling of α2,3-Linked Sialic Acid Residues (2019)
  • Author(s): Hanamatsu Hisatoshi、Nishikaze Takashi、Tsumoto Hiroki、Ogawa Koji、Kobayashi Takashi、Yokota Ikuko、Morikawa Kenichi、Suda Goki、Sho Takuya、Nakai Masato、Miura Nobuaki、Higashino Kenichi、Sekiya Sadanori、Iwamoto Shinichi、Miura Yuri、Furukawa Jun-ichi、Tanaka Koichi、Sakamoto Naoya
  • Journal Title: Analytical Chemistry Volume: 91 Issue: 21 Pages: 13343-13348
  • DOI: 10.1021/acs.analchem.9b03617
  • Peer Reviewed
• [Presentation] シアル酸結合様式特異的な化学修飾による構造異性体識別法の開発 (2021)
  • Author(s): 花松久寿
  • Organizer: 第40回日本糖質学会年会
• [Presentation] 安定同位体を用いたSALSA法による比較質量分析法の開発 (2020)
  • Author(s): 花松久寿, 西風隆司, 津元 裕樹, 小川浩司, 横田育子, 森川賢一, 須田剛生, 荘拓也, 中井正人, 三浦信明, 関谷禎規, 岩本慎一, 三浦ゆり, 田中 耕一, 坂本直哉, 古川潤一
  • Organizer: 第39回日本糖質学会年会
• [Presentation] ケミカルグライコミクスを用いた加齢による糖鎖変化の包括的解析 (2020)
  • Author(s): 花松久寿, 古川潤一
  • Organizer: 第93回日本生化学会大会
  • Invited
• [Patent(Industrial Property Rights)] 分析用試料の作成方法 (2019)
  • Inventor(s): 花松 久寿、西風 隆司、横田 育子、古川 潤一
  • Industrial Property Rights Holder: 花松 久寿、西風 隆司、横田 育子、古川 潤一
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-130188
  • Filing Date: 2019