Background: Human coagulation factor IX (hFIX) is a glycoprotein with two N-glycosylation sites at the activation peptide. Since the activation peptide is removed in mature hFIX, the exact role of N-glycosylation is unclear. To investigate the role of N-glycosylation in the secretion and activity of hFIX, we inhibited N-glycosylation by tunicamycin in the stable Human Embryonic Kidney (HEK)coagulation Factor IX (FIX) cells. Materials and Methods: After the treatment of stable FIX-expressing HEK cells in the presence or absence of tunicamycin, the expression and activity of the recombinant FIX (rFIX) were determined in culture medium and cell lysate with enzyme-linked immunosorbent assay and clotting test, respectively. Results: Based on the data analysis, total concentrations of FIX in stable HEK-FIX was the same in the media with and without tunicamycin. But throughout the post-induction period, the intracellular and secreted levels of FIX in tunicamycin-treated HEK-FIX cells increased and decreased, respectively, compared with those of control HEK-FIX cells, though the results were not significant. These results indicate that disrupting the synthetic process may slightly reduce the FIX levels secreted in HEK-FIX cells. Conclusion: Although glycosylation plays a vital role in the folding and secretion of the proteins, it does not affect the secretion of FIX. Besides, the N-glycosylation of the produced FIX failed to play a significant role in its activity.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.