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Chemoenzymatic Approach for the Proteomics Analysis of Mucin-Type Core-1 O-Glycosylation in Human Serum

文献类型：期刊论文

作者You, Xin2,3; Yao, Yating2,3; Mao, Jiawei2,3; Qin, Hongqiang2,3; Liang, Xinmiao2,3; Wang, Liming1; Ye, Mingliang2,3
刊名ANALYTICAL CHEMISTRY
出版日期2018-11-06
卷号90期号:21页码:12714-12722
ISSN号0003-2700
DOI10.1021/acs.analchem.8b02993
通讯作者Ye, Mingliang(mingliang@dicp.ac.cn)
英文摘要Human serum is a complex body fluid that contains various N-linked and O-linked glycoproteins. Compared with N-linked glycoproteins, the serum O-linked glycoproteins are not well-studied due to their high heterogeneity and their low abundance. Herein, we presented a novel chemoenzymatic method to analyze core-1 type of O-GalNAcylation in human serum. In this approach, the tryptic digest of serum was first subjected to PNGase F treatment to release the N-glycan and was then treated with strong acid to release sialic acid residues from mucin-type O-glycans. In this way, the internal Gal/GalNAc residues were exposed and were oxidized by the galactose oxidase to carry the aldehyde groups. The oxidized O-GalNAcylated peptides were then captured by hydrazide beads and eluted with methoxylamine for LC-MS/MS analysis. The de-N-deglycosylation decreased the abundance of N-glycopeptides, the desialylation simplified the O-glycans and the enzymatic oxidization conferred the enrichment specificity. We have demonstrated that this method was fitted to analyze O-GalNAcylated peptides with high confidence. This method was applied to analyze human serum, which resulted in the identification of 59 O-GalNAc modified peptide sequences corresponding to 38 glycoproteins from 50 mu L of serum. This method is expected to have broad applications in the analysis of O-glycoproteome.
WOS关键词INTERACTION LIQUID-CHROMATOGRAPHY ; LARGE GENE LISTS ; MASS-SPECTROMETRY ; ENRICHMENT ; IDENTIFICATION ; GLYCOPEPTIDES ; PROTEINS ; STRATEGY ; CELLS ; GLYCOPROTEOME
资助项目China State Key Basic Research Program Grants[2018YFC0910302] ; China State Key Basic Research Program Grants[2016YFA0501402] ; China State Key Basic Research Program Grants[2017YFA0505003] ; National Natural Science Foundation of China[21535008] ; National Natural Science Foundation of China[81430072] ; National Natural Science Foundation of China[21775146] ; National Natural Science Foundation of China[81272368] ; National Natural Science Foundation of China[81471755] ; Innovation Program of Science and Research from the DICP, CAS[DICP TMSR201601] ; National Science Fund of China for Distinguished Young Scholars[21525524]
WOS研究方向Chemistry
语种英语
出版者AMER CHEMICAL SOC
WOS记录号WOS:000449722500054
资助机构China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Innovation Program of Science and Research from the DICP, CAS ; Innovation Program of Science and Research from the DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Innovation Program of Science and Research from the DICP, CAS ; Innovation Program of Science and Research from the DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Innovation Program of Science and Research from the DICP, CAS ; Innovation Program of Science and Research from the DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars ; China State Key Basic Research Program Grants ; China State Key Basic Research Program Grants ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; Innovation Program of Science and Research from the DICP, CAS ; Innovation Program of Science and Research from the DICP, CAS ; National Science Fund of China for Distinguished Young Scholars ; National Science Fund of China for Distinguished Young Scholars
源URL[http://cas-ir.dicp.ac.cn/handle/321008/166628]  
专题大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者Ye, Mingliang
作者单位1.Dalian Med Univ, Hosp 2, Dept Surg, Div Hepatobiliary & Pancreat Surg, Dalian 116023, Liaoning, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714		 You, Xin,Yao, Yating,Mao, Jiawei,et al. Chemoenzymatic Approach for the Proteomics Analysis of Mucin-Type Core-1 O-Glycosylation in Human Serum[J]. ANALYTICAL CHEMISTRY,2018,90(21):12714-12722.
APA You, Xin.,Yao, Yating.,Mao, Jiawei.,Qin, Hongqiang.,Liang, Xinmiao.,...&Ye, Mingliang.(2018).Chemoenzymatic Approach for the Proteomics Analysis of Mucin-Type Core-1 O-Glycosylation in Human Serum.ANALYTICAL CHEMISTRY,90(21),12714-12722.
MLA You, Xin,et al."Chemoenzymatic Approach for the Proteomics Analysis of Mucin-Type Core-1 O-Glycosylation in Human Serum".ANALYTICAL CHEMISTRY 90.21(2018):12714-12722.

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来源：大连化学物理研究所

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