A new approach for mapping sialylated N-glycosites in serum proteomes.
A new approach for proteome-wide analysis of sialylated N-glycopeptides based on the diagonal chromatographic COFRADIC technology is presented here. The use of alpha(2-3,6,8,9) neuraminidase is central to isolate sialylated N-glycopeptides out of a complex peptide mixture. Two different COFRADIC techniques are introduced here, either without or with post-metabolic oxygen-18 labeling (direct versus indirect sorting), and when applied to immuno-depleted mouse serum, we herewith identified 93 sialylated glycosylation sites in 53 serum proteins.
Ghesquière
Bart
B
Department of Medical Protein Research, VIB, B-9000 Ghent, Belgium.
Buyl
Lien
L
Demol
Hans
H
Van Damme
Jozef
J
Staes
An
A
Timmerman
Evy
E
Vandekerckhove
Joël
J
Gevaert
Kris
K
D000595:Amino Acid Sequence; D000818:Animals; D001798:Blood Proteins / Q000737:chemistry*; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006020:Glycopeptides / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000737:chemistry; D010103:Oxygen Isotopes / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D012794:Sialic Acids / Q000737:chemistry*; D012984:Software
D001798:Blood Proteins; D006020:Glycopeptides; D010103:Oxygen Isotopes; D012794:Sialic Acids; D009439:Neuraminidase