Comprehensive mapping of protein N-glycosylation in human liver by combining hydrophilic interaction chromatography and hydrazide chemistry.
Although glycoproteomics is greatly developed in recent years, our knowledge about N-glycoproteome of human tissues is still very limited. In this study, we comprehensively mapped the N-glycosylation sites of human liver by combining click maltose-hydrophilic interaction chromatography (HILIC) and the improved hydrazide chemistry. The specificity could be as high as 90% for hydrazide chemistry and 80% for HILIC. Altogether, we identified 14,480 N-glycopeptides matched with N-!P-[S|T|C] sequence motif from human liver, corresponding to 2210 N-glycoproteins and 4783 N-glycosylation sites. These N-glycoproteins are widely involved into different types of biological processes, such as hepatic stellate cell activation and acute phase response of human liver, which all highly associate with the progression of liver diseases. Moreover, the exact N-glycosylation sites of some key-regulating proteins within different human liver physiological processes were also obtained, such as E-cadherin, transforming growth factor beta receptor and 29 members of G protein coupled receptors family.
Key Lab of Separation Science for Analytical Chemistry, National Chromatographic Research and Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences , 457 Zhongshan Road, Dalian 116023, China.
Jun
J
Zhu
Zhen
Z
Sun
Kai
K
Cheng
Rui
R
Chen
Mingliang
M
Ye
Bo
B
Xu
Deguang
D
Sun
Liming
L
Wang
Jing
J
Liu
Fangjun
F
Wang
Hanfa
H
Zou
D020816:Amino Acid Motifs; D015820:Cadherins / Q000032:analysis / Q000737:chemistry; D002845:Chromatography / Q000379:methods; D005260:Female; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D006834:Hydrazines / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D008099:Liver / Q000737:chemistry* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D040901:Proteomics; D043562:Receptors, G-Protein-Coupled / Q000032:analysis / Q000737:chemistry; D018125:Receptors, Transforming Growth Factor beta / Q000032:analysis / Q000737:chemistry
D015820:Cadherins; D006023:Glycoproteins; D006834:Hydrazines; D043562:Receptors, G-Protein-Coupled; D018125:Receptors, Transforming Growth Factor beta