N-Glycan structure annotation of glycopeptides using a linearized glycan structure database (GlyDB).
While glycoproteins are abundant in nature, and changes in glycosylation occur in cancer and other diseases, glycoprotein characterization remains a challenge due to the structural complexity of the biopolymers. This paper presents a general strategy, termed GlyDB, for glycan structure annotation of N-linked glycopeptides from tandem mass spectra in the LC-MS analysis of proteolytic digests of glycoproteins. The GlyDB approach takes advantage of low-energy collision-induced dissociation of N-linked glycopeptides that preferentially cleaves the glycosidic bonds while the peptide backbone remains intact. A theoretical glycan structure database derived from biosynthetic rules for N-linked glycans was constructed employing a novel representation of branched glycan structures consisting of multiple linear sequences. The commonly used peptide identification program, Sequest, could then be utilized to assign experimental tandem mass spectra to individual glycoforms. Analysis of synthetic glycopeptides and well-characterized glycoproteins demonstrate that the GlyDB approach can be a useful tool for annotation of glycan structures and for selection of a limited number of potential glycan structure candidates for targeted validation.
Ren
Jian Min
JM
Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, Massachusetts 02115, USA.
Rejtar
Tomas
T
Li
Lingyun
L
Karger
Barry L
BL
D002240:Carbohydrate Sequence; D030562:Databases, Protein; D006020:Glycopeptides / Q000737:chemistry*; D006031:Glycosylation; D008962:Models, Theoretical; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
D006020:Glycopeptides; D011134:Polysaccharides