Title :
Large-scale intact
glycopeptide identification by Mascot database search
Abstract :
- Workflows capable of determining glycopeptides in large-scale are missing in the field of glycoproteomics
- We present an approach for automated annotation of intact glycopeptide mass spectra
- The steps in adopting the Mascot search engine for intact glycopeptide analysis included: (i) assigning one letter codes for monosaccharides, (ii) linearizing glycan sequences and (iii) preparing custom glycoprotein databases
- Automated annotation of both N- and O-linked glycopeptides was proven using standard glycoproteins
- In a large-scale study, a total of 257 glycoproteins containing 970 unique glycosylation sites and 3447 non-redundant N-linked glycopeptide variants were identified in 24 serum samples
- Thus, a single tool was developed that collectively allows the (i) elucidation of N- and O-linked glycopeptide spectra, (ii) matching glycopeptides to known protein sequences , and (iii) high-throughput, batch-wise analysis of large-scale glycoproteomics data sets