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Analysis of fucosylation in liver-secreted N-glycoproteins from human hepatocellular carcinoma plasma using liquid chromatography with tandem mass spectrometry

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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.

A global map of fucosylated and non-fucosylated glycopeptides from 13 liver-secreted glycoproteins in hepatocellular carcinoma plasma

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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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    Authors and Affiliations

    1. Biomedical Omics Group, Korea Basic Science Institute, 162 YeonGuDanji-Ro, Ochang-eup, Cheongju, Chungbuk, 28119, Republic of Korea

      Eun Sun Ji, Heeyoun Hwang, Gun Wook Park, Ju Yeon Lee, Hyun Kyoung Lee, Na Young Choi, Hoi Keun Jeong, Kwang Hoe Kim, Jin Young Kim & Jong Shin Yoo

    2. Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, 305-764, Republic of Korea

      Gun Wook Park, Hyun Kyoung Lee, Na Young Choi, Hoi Keun Jeong, Kwang Hoe Kim & Jong Shin Yoo

    3. Department of Chemistry, Hannam University, Daejeon, 306-791, Republic of Korea

      Seungho Lee

    4. Department of Biomedical Science, Cheongju University, Cheongju, 28503, Republic of Korea

      Yeong Hee Ahn

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    1. Eun Sun Ji
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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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