A novel mass spectrometric procedure for the rapid determination of the types of carbohydrate chains present in glycoproteins: application to alpha-galactosidase I from Vicia faba seeds.
A fast atom bombardment mass spectrometric protocol has been developed to determine the type of oligosaccharide chain present in glycoproteins. The procedure is based on acetolysis of the intact glycoconjugate, extraction of the peracetylated carbohydrate fragments and analysis by fast atom bombardment mass spectrometry. The molecular ions present in the FAB spectra uniquely define the composition of the oligosaccharides with respect to hexose, aminohexose and sialic acid content. High mannose oligosaccharides yield a series of peracetylated hexose oligomers whereas complex-type oligosaccharides afford a series of N-acetyl-lactosamine containing species. Fucosylation is usually not detected but sialylated oligosaccharides are readily identified and the type of sialic acid is also defined. The method has been tested on three glycoproteins of known structure - fetuin, ribonuclease B and erythrocyte Band 3 - and on a glycoprotein of unknown structure - alpha-galactosidase I, an enzyme lectin from Vicia faba. The latter is shown to contain high mannose carbohydrate chains.
S
S
Naik
J E
JE
Oates
A
A
Dell
G W
GW
Taylor
P M
PM
Dey
J B
JB
Pridham
D001457:Anion Exchange Protein 1, Erythrocyte / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis*; D005696:Galactosidases / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D012260:Ribonucleases / Q000032:analysis; D012639:Seeds / Q000032:analysis*; D013997:Time Factors; D000509:alpha-Fetoproteins / Q000032:analysis; D000519:alpha-Galactosidase / Q000032:analysis*
D001457:Anion Exchange Protein 1, Erythrocyte; D002241:Carbohydrates; D000509:alpha-Fetoproteins; D012260:Ribonucleases; C029600:ribonuclease B; D005696:Galactosidases; D000519:alpha-Galactosidase