Identification of O-glycoproteins by Glycoproteomics based on BEP procedure
Project/Area Number |
16K05804
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Analytical chemistry
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Research Institution | Hokkaido University |
Principal Investigator |
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Project Period (FY) |
2016-04-01 – 2019-03-31
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Project Status |
Completed (Fiscal Year 2018)
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Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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Keywords | 糖タンパク質 / グライコミクス / β脱離反応 / マイケル付加 / O-結合型糖鎖 / BEP法 / プロテオミクス |
Outline of Final Research Achievements |
Serine (Ser) and threonine (Thr) residues are modified by the O-GalNAc type, typical of mucin. The characterization of O-glycans has remained particularly challenging because no analogous endoglycosidase is known. We investigated the BEP method for the identification of structural isomer and O-glycosylation sites of proteins/peptides. We developed that aminolysis-SALSA could successfully distinguish sialylated N- and GSL-glycan isomers by mass spectrometry. This method could be applied to the analysis of sialylated O-glycopeptides. In addition, we investigated the BEP procedure focused on Michael addition. In this study, it was revealed that only serine residues act as Michael acceptor and preform selective Michael addition. Furthermore, we made it clear that the efficiency of cleavage of O-glycan differed between Ser and Thr residue.
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Academic Significance and Societal Importance of the Research Achievements |
本研究でのシアル酸の構造上の識別法、スレオニン残基修飾糖鎖の切断法、そしてマイケル付加反応の効率についての研究により、これまでに詳細な情報がなく、本研究の成果が糖タンパク質に関連する診断マーカー探索研究や糖鎖付加部位さらにはキャリアタンパク質を同定するために重要な情報取得に繋がる。
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Report
(4 results)
Research Products
(30 results)
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[Presentation] BEP法を基盤とした糖鎖および糖鎖結合部位の同定法2017
Author(s)
岡田 和恵, 横田 育子, 花松 久寿, 大西 俊介, 湯山 耕平, 酒井 祥太, 伊東 信, 五十嵐 靖之, 坂本 直哉, 篠原 康郎, 古川 潤一
Organizer
第36回 日本糖質学会
Related Report
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[Book] 実験医学2017
Author(s)
古川 潤一,岩崎 倫政,篠原 康郎
Total Pages
7
Publisher
羊土社
Related Report