( arily of sialic acid galactose ...)Structural Analysis of Glycosylated Peptides in Complex Mixtures with ,,, Ion Trap MSn

arily of sialic acid, galactose and N-acetylglucosamine.

Figure 3 designates the expected mass-to-charge ratios for the possible forms of the glycosylated tryptic peptide T45 from rt-PA. This glycoform was found to have a typical structure including one sialic acid, two galactoses and two N-acetylglucosamines (GlcN) (labeled as structure 211). Figure 2. Sequence of rt-PA. Observed peptides labeled in red. Blue highlighted peptides correspond to glycoforms identified after possible glycoform structures were manually added to the database. Small peptides in black, representing 10% of the amino acids, were not observed.

Figure 3. Typical theoretical oligosaccharide sequences for rt-PA peptide T45. Mass-to-charge ratios and charge states for the heterogeneous structures are listed in parentheses. The unglycosylated peptide was determined to have a molecular weight of 1131 amu.

Figure 4. Top panel Base peak ion chromatogram of tryptic peptides from rt-PA.
Bottom panel Full-scan mass spectrum of the 211-T45 glycoform eluting at 15.57 minutes.

Figure 5. MS/MS on the 211-T45 ion at m/z 1064. The glycopeptide fragmented into many different glycoforms. The 210-G, GlcN ion m/z 1267.6 (+2) was selected for MS³ analysis.
Theoretical m/z values for all possible cleavage products of the glycoform were added to the database to enable TurboSEQUEST to automatically match glycopeptide structures. From these, TurboSEQUEST was
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