Chromatographic investigation of the glycosylation pattern of alpha-1-acid glycoprotein secreted by the HepG2 cell line; a putative model for inflammation?
In certain pathophysiological conditions, such as rheumatoid arthritis, there are alterations in the glycosylation pattern of the acute phase protein, alpha-1-acid glycoprotein (AGP). These changes are likely to be functionally significant, however, verification of the latter role requires a system which reflects in vivo glycosylation changes in AGP and also produces sufficient quantities of the protein for further study. The human hepatoma cell line HepG2 is documented as displaying a shift in the glycosylation pattern of glycoproteins from normal state to acute phase after stimulation with inflammatory mediators. We have isolated AGP from the culture medium of HepG2 cells both before and after stimulation with a cytokine preparation and analysed the glycosylation pattern of each preparation, after enzymatic release, by high pH anion-exchange chromatography. Before stimulation, the glycosylated population was similar to a profile of AGP isolated from normal plasma; however, cytokine stimulation resulted in a shift to a profile which was consistent with that of AGP from a rheumatoid arthritis sufferer. Thus a HepG2 cell culture system is capable of being a crude model of the changes in glycosylation of acute phase proteins although it has a tendency to produce oligosaccharide chains which are not fully sialylated.
Elliott
H G
HG
Department of Pharmaceutical Sciences, University of Strathelyde, Glasgow, UK.
Elliott
M A
MA
Watson
J
J
Steele
L
L
Smith
K D
KD
D000818:Animals; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D016207:Cytokines / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D009961:Orosomucoid / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D016207:Cytokines; D009961:Orosomucoid; D006026:Glycoside Hydrolases