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Detergent-Assisted Glycoprotein Capture: A Versatile Tool for In-Depth N-Glycoproteome Analysis
Chen, Rui1,2; Zou, Hanfa3; Figeys, Daniel1,2
KeywordDetergent Glycoprotein Capture Membrane Serum N-glycoproteome
Source PublicationJOURNAL OF PROTEOME RESEARCH
2016-06-01
ISSN1535-3893
DOI10.1021/acs.jproteome.6b00056
Volume15Issue:6Pages:2080-2086
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordHYDROPHILIC INTERACTION CHROMATOGRAPHY ; PROTEIN GLYCOSYLATION ; HYDRAZIDE CHEMISTRY ; MASS-SPECTROMETRY ; PLASMA PROTEOME ; IDENTIFICATION ; CANCER ; LIVER ; DISINTEGRIN ; PROTEOLYSIS
AbstractLarge-scale N-glycoproteome studies have been hindered by poor solubility of hydrophobic membrane proteins and the complexity of proteome samples. Herein, we developed a detergent-assisted glycoprotein capture method to reduce these issues by conducting hydrazide chemistry-based glycoprotein capture in the presence of strong detergents such as sodium dodecyl sulfate and Triton X-100. The strong detergents helped to solubilize hydrophobic membrane proteins and then increased the access of hydrazide groups to oxidized glycoproteins, thus increasing the coverage of the N-glycoproteome. Compared with the conventional glycopeptide capture method, the detergent-assisted glycoprotein capture approach nearly doubled the number of N-glycosylation sites identified from HEK 293T cells with improved specificity. Application of this approach in the larger scale N-glycoproteomics analysis of the HEK 293T cell membrane led to the identification of 2253 unique N-glycosites from 953 proteins. Furthermore, the application of this approach to human serum resulted in the identification of 850 N-glycosylation sites without any immunodepletion or fractionation. Overall, the detergent-assisted glycoprotein capture method simplified the capture process, and it increased the number of sites observed on both hydrophobic membrane proteins and hydrophilic secreted proteins.
Language英语
WOS IDWOS:000377319200030
PublisherAMER CHEMICAL SOC
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/170613
Collection中国科学院大连化学物理研究所
Corresponding AuthorFigeys, Daniel
Affiliation1.Univ Ottawa, Dept Biochem Microbiol & Immunol, Ottawa Inst Syst Biol, 451 Smyth Rd, Ottawa, ON K1H 8M5, Canada
2.Univ Ottawa, Dept Chem & Biomol Sci, 451 Smyth Rd, Ottawa, ON K1H 8M5, Canada
3.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
Chen, Rui,Zou, Hanfa,Figeys, Daniel. Detergent-Assisted Glycoprotein Capture: A Versatile Tool for In-Depth N-Glycoproteome Analysis[J]. JOURNAL OF PROTEOME RESEARCH,2016,15(6):2080-2086.
APA Chen, Rui,Zou, Hanfa,&Figeys, Daniel.(2016).Detergent-Assisted Glycoprotein Capture: A Versatile Tool for In-Depth N-Glycoproteome Analysis.JOURNAL OF PROTEOME RESEARCH,15(6),2080-2086.
MLA Chen, Rui,et al."Detergent-Assisted Glycoprotein Capture: A Versatile Tool for In-Depth N-Glycoproteome Analysis".JOURNAL OF PROTEOME RESEARCH 15.6(2016):2080-2086.
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