Abstract
Fucosylation of N-glycoproteins has been implicated in various diseases, such as hepatocellular carcinoma (HCC). However, few studies have performed site-specific analysis of fucosylation in liver-secreted proteins. In this study, we characterized the fucosylation patterns of liver-secreted proteins in HCC plasma using a workflow to identify site-specific N-glycoproteins, where characteristic B- and/or Y-ion series with and without fucose in collision-induced dissociation were used in tandem mass spectrometry. In total, 71 fucosylated N-glycopeptides from 13 major liver-secreted proteins in human plasma were globally identified by LC-MS/MS. Additionally, 37 fucosylated N-glycopeptides were newly identified from nine liver-secreted proteins, including alpha-1-antichymotrypsin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, ceruloplasmin, alpha-1-acid glycoprotein 1/2, alpha-2-macroglobulin, serotransferrin, and beta-2-glycoprotein 1. Of the fucosylated N-glycopeptides, bi- and tri-antennary glycoforms were the most common ones identified in liver-secreted proteins from HCC plasma. Therefore, we suggest that this analytical method is effective for characterizing fucosylation in liver-secreted proteins.
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Abbreviations
- A2M:
-
Alpha-2-macroglobulin
- AACT:
-
Alpha-1-antichymotrypsin
- AAT:
-
Alpha-1-antitrypsin
- AGP1:
-
Alpha-1-acid glycoprotein 1
- AGP2:
-
Alpha-1-acid glycoprotein 2
- APOH:
-
Beta-2-glycoprotein 1
- CID:
-
Collision-induced dissociation
- CP:
-
Ceruloplasmin
- FETUA:
-
Fetuin A
- Fuc:
-
Fucose
- HCC:
-
Hepatocellular carcinoma
- HCD:
-
Higher-energy collision dissociation
- Hex:
-
Hexose
- HexNAc:
-
N-Acetyl hexosamine
- HILIC:
-
Hydrophilic interaction liquid chromatography
- HP:
-
Haptoglobin
- HPX:
-
Hemopexin
- KNG1:
-
Kininogen-1
- MS:
-
Mass spectrometry
- N:
-
Asparagine
- Sia:
-
Sialic acid
- TF:
-
Serotransferrin
- VTN:
-
Vitronectin
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Acknowledgments
This research was supported by the National Research Council of Science and Technology (NTM2371511, the Creative Allied Project (CAP)), the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI); it was funded by the Ministry of Health &Welfare, Republic of Korea (grant number: HI13C2098), and by the Ministry of Trade, Industry & Energy (MOTIE, 1415139249) through the Osong Academy–Industry Convergence (BAIO) fostering project.
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This study was performed in accordance with ethical standards. Plasma samples were obtained from voluntary blood donors with informed consent and approval in accordance with IRB guidelines from Yonsei University College of Medicine (Seoul, Korea).
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Eun Sun Ji and Heeyoun Hwang contributed equally to this work.
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Ji, E.S., Hwang, H., Park, G.W. et al. Analysis of fucosylation in liver-secreted N-glycoproteins from human hepatocellular carcinoma plasma using liquid chromatography with tandem mass spectrometry. Anal Bioanal Chem 408, 7761–7774 (2016). https://doi.org/10.1007/s00216-016-9878-0
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DOI: https://doi.org/10.1007/s00216-016-9878-0