Title : Analysis of site-specific glycosylation in recombinant human
follistatin expressed in Chinese hamster ovary cells
Abstract :
- Follistatin (FS) FS ), a glycoprotein , plays an important role in cell growth and differentiation through the neutralization of the biological activities of activins
- In this study, we analyzed the glycosylation of recombinant human FS ( rhFS ) produced in Chinese hamster ovary cells
- The results of SDS-PAGE and MALDI-TOF MS revealed the presence of both non-glycosylated and glycosylated forms
- FS contains two potential N-glycosylation sites, Asn95 and Asn259
- Using mass spectrometric peptide/glycopeptide mapping and precursor-ion scanning, we found that both N-glycosylation sites were partially glycosylated
- Monosaccharide com position analyses suggested the linkages of fucosylated bi- and triantennary complex-type oligosaccharides on rhFS
- This finding was supported by mass spectrometric oligosaccharide profiling , in which the m/z values and elution times of some of the oligosaccharides from rhFS were in good agreement with those of standard oligosaccharides
- Site-specific glycosylation was deduced on the basis of the mass spectra of the glycopeptides
- It was suggested that biantennary oligosaccharides are major oligosaccharides located at both Asn95 and Asn259 , whereas the triantennary structures are present mainly at Asn95