Comparative glycoproteomics based on lectins affinity capture of N-linked glycoproteins from human Chang liver cells and MHCC97-H cells.
We present here an effective technique for the large-scale separation and identification of N-linked glycoproteins from Chang liver cells, the human normal liver cells. To enrich N-linked glycoproteins from the whole cells, a procedure containing the lysis of human liver cells, the solubilization of total proteins, lectin affinity chromatography including Concanavalin A and wheat germ agglutinin was established. Furthermore, captured N-linked glycoproteins were separated by 2-DE, and identified by MS and database searching. Finally, we found 63 N-glycoproteins in Chang liver cells. In addition, using the above method, we identified 7 remarkably up-regulated glycoproteins from MHCC97-H cells, highly metastatic liver cancer cells, compared to Chang liver cells. These up-regulated glycoproteins were associated with liver cancer and might be used as biomarkers for tumor diagnosis. Results showed that we established a high-throughput proteomic analysis for separating N-linked glycoproteins from human liver cells. This strategy greatly improved the glycoprotein analysis method associated with proteome-wide glycosylation changes related to liver cancer. Our work was part of the HUPO Human Liver Proteome Project (HLPP) studies and was supported by CHINA HUPO.
Xu
Zhibin
Z
Glycoconjugate Key Laboratory, Ministry of Health, Department of Biochemistry and Molecular Biology, Fudan University, Shanghai, PR China.
Zhou
Xinwen
X
Lu
Haojie
H
Wu
Nan
N
Zhao
Hongbo
H
Zhang
Lineng
L
Zhang
Wen
W
Liang
Yu Long
YL
Wang
Liying
L
Liu
Yinkun
Y
Yang
Pengyuan
P
Zha
Xiliang
X
D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D022781:Hepatocytes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015854:Up-Regulation; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
D006023:Glycoproteins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A