Isolation and identification of sialylated glycopeptides from bovine alpha1-acid glycoprotein by off-line capillary electrophoresis MALDI-TOF mass spectrometry.
Sialylated glycopeptides contained in liquid chromatographic fractions of bovine alpha1-glycoprotein tryptic digests were isolated from asialo peptides using capillary electrophoresis (CE). CE effluents were deposited directly onto a metallic target and analyzed using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. This method allowed the characterization of four N-glycosylation sites in the glycoprotein, and each site was observed as a set of sialylated peptide glycoforms. Tandem mass spectrometry was used to confirm peptide sequences and glycan content in glycoforms. The CE method developed for this study resulted in a very clear separation of the sialylated from the asialo content of glycoprotein digests and proved very useful in the determination of the nature and location of sialylated glycans along the protein chain. This article is the first report describing the use of on-target CE fraction collection using a MALDI removable sample concentrator.
Snovida
Sergei I
SI
Department of Chemistry, University of Manitoba, Winnipeg, Manitoba, Canada R3T 2N2.
Chen
Vincent C
VC
Krokhin
Oleg
O
Perreault
Hélène
H
D000818:Animals; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry* / Q000302:isolation & purification; D009961:Orosomucoid / Q000737:chemistry*; D012794:Sialic Acids / Q000737:chemistry; D012795:Sialoglycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D018629:Spectrometry, Mass, Secondary Ion
D006020:Glycopeptides; D009961:Orosomucoid; D012794:Sialic Acids; D012795:Sialoglycoproteins