Glycosylation sites isolation

Human biotinidase is a monomeric glycoprotein with a molecular mass of 76 kDa when analysed by sodium dodecylsulphate-polyacrylamide gel electrophoresis [7]. The cDNA for human biotinidase was isolated and sequenced in 1994 by Cole and coworkers [6]. The deduced protein contains 502 amino acids, has a molecular mass of 57 kDa and contains 6 potential N-linked glycosylation sites. The biotinidase gene... 

FIG. 7. Identification of four glycosylation sites of the isolated MUC4 derivative by ECD-MS/MS analysis. (A) Results of the spectral matching analysis using 30 possible structures. (B) the top four structures predicted by in silico calculation and the spectral coverage (%) when compared with die real experimental spectrum. This figure is adapted from ref. 77. (Continued)... 

After isolation and identification of the peptides containing these glycosylation sites, high pH anion exchange chromatography, matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, and electrospray ionization mass spectrometry (ES-MS) were utilized to evaluate carbohydrate microheterogeneity. 

Fig. 2. Amino acid sequences of L-PSGPs isolated from fertilized eggs of salmonid fish species where indicates the 0-glycosylation sites. H-PSGPs isolated from unfertilized eggs are polymerized forms of L-PSGP.

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