TY - JOUR
T1 - Site-Specific Profiling of Serum Glycoproteins Using N-Linked Glycan and Glycosite Analysis Revealing Atypical N-Glycosylation Sites on Albumin and α-1B-Glycoprotein
AU - Sun, Shisheng
AU - Hu, Yingwei
AU - Jia, Li
AU - Eshghi, Shadi Toghi
AU - Liu, Yang
AU - Shah, Punit
AU - Zhang, Hui
N1 - Funding Information:
This work was supported by the National Institutes of Health, National Cancer Institute, the Early Detection Research Network (EDRN; Grant U01CA152813), the Clinical Proteomic Tumor Analysis Consortium (CPTAC; Grant U24CA210985), National Heart Lung and Blood Institute, Programs of Excellence in Glycosciences (PEG; Grant P01HL107153), and the National Institute of Allergy and Infectious Diseases (Grant R21AI122382). The work was also supported by National Natural Science Foundation of China (Grant Nos. 81773180 and 21705127).
Publisher Copyright:
© 2018 American Chemical Society.
PY - 2018/5/15
Y1 - 2018/5/15
N2 - Most serum proteins are N-linked glycosylated, and therefore the glycoproteomic profiling of serum is essential for characterization of serum proteins. In this study, we profiled serum N-glycoproteome by our recently developed N-glycoproteomic method using solid-phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) coupled with LC-MS/MS and site-specific glycosylation analysis using GPQuest software. Our data indicated that half of identified N-glycosites were modified by at least two glycans, with a majority of them being sialylated. Specifically, 3/4 of glycosites were modified by biantennary N-glycans and 1/3 of glycosites were modified by triantennary sialylated N-glycans. In addition, two novel atypical glycosites (with N-X-V motif) were identified and validated from albumin and α-1B-glycoprotein. The widespread presence of these two glycosites among individuals was further confirmed by individual serum analyses.
AB - Most serum proteins are N-linked glycosylated, and therefore the glycoproteomic profiling of serum is essential for characterization of serum proteins. In this study, we profiled serum N-glycoproteome by our recently developed N-glycoproteomic method using solid-phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) coupled with LC-MS/MS and site-specific glycosylation analysis using GPQuest software. Our data indicated that half of identified N-glycosites were modified by at least two glycans, with a majority of them being sialylated. Specifically, 3/4 of glycosites were modified by biantennary N-glycans and 1/3 of glycosites were modified by triantennary sialylated N-glycans. In addition, two novel atypical glycosites (with N-X-V motif) were identified and validated from albumin and α-1B-glycoprotein. The widespread presence of these two glycosites among individuals was further confirmed by individual serum analyses.
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U2 - 10.1021/acs.analchem.8b01051
DO - 10.1021/acs.analchem.8b01051
M3 - Article
C2 - 29671580
AN - SCOPUS:85046488886
VL - 90
SP - 6292
EP - 6299
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 10
ER -