Targeted proteo-glycomics analysis of Sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line.
Sialyl Lewis X (SLEX) antigen, Neu5Acalpha2-3Galbeta1-4 (Fucalpha1-3) GlcNAc-R, plays important roles in cell-to-cell interaction: for example, the E- and P-selectin-mediated influx of SLEX expressing leukocytes into inflamed areas. A human hepatocellular carcinoma cell line, HepG2 cells, was highly expressed SLEX on secreted glycoproteins and cell surface, in contrast with HuH-7 cells. We identified SLEX expressing glycoproteins in HepG2 cultured medium by two-dimensional polyacrylamide gel electrophoresis, followed by in gel digestion and peptide mass fingerprint using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS), including transferrin, alpha1-antitrypsin, alpha2-HS glycoprotein and beta-glycoprotein. We analyzed N-glycans of these glycoproteins by MALDI-TOFMS in combination with exoglycosidase digestion; our results indicate increases in poly-fucosylated and high-branched N-glycans. High alpha1,3-fucosylation in glycoproteins would be caused by increased expression of alpha1,3-fucosyltransferase activities in HepG2 cells.
Higai
Koji
K
Department of Clinical Chemistry, School of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba 274-8510, Japan. koji@phar.toho-u.ac.jp
Shibukawa
Kumiko
K
Muto
Satoshi
S
Matsumoto
Kojiro
K
D006528:Carcinoma, Hepatocellular / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D016256:Lewis X Antigen / Q000737:chemistry / Q000276:immunology*; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology*; D040901:Proteomics; D000080506:Sialyl Lewis X Antigen; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014407:Tumor Cells, Cultured
D006023:Glycoproteins; D016256:Lewis X Antigen; D009844:Oligosaccharides; D000080506:Sialyl Lewis X Antigen