Comparative glycoproteomics based on lectins affinity capture of N-linked glycoproteins from human Chang liver cells and MHCC97-H cells

• Published in: Proteomics (Weinheim) Vol. 7; no. 14; pp. 2358 - 2370
• Main Authors: Xu, Zhibin, Zhou, Xinwen, Lu, Haojie, Wu, Nan, Zhao, Hongbo, Zhang, Lineng, Zhang, Wen, Liang, Yu Long, Wang, Liying, Liu, Yinkun, Yang, Pengyuan, Zha, Xiliang
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.07.2007; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Cells, Cultured; Chromatography, Affinity - methods; Concanavalin A - metabolism; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Glycoprotein; Glycoproteins - analysis; Glycoproteins - chemistry; Glycoproteins - metabolism; Glycoproteomics; Hepatocytes - chemistry; Hepatocytes - metabolism; Humans; Lectin; Liver; Miscellaneous; Proteins; Proteomics - methods; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Up-Regulation; Wheat Germ Agglutinins - metabolism; Human; Lectin; Glycoproteomics; Liver; Glycoprotein; Proteomics
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200600041

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.