Electrospray ionization-ion trap mass spectrometry for structural analysis of complex N-linked glycoprotein oligosaccharides.
Electrospray ionization-ion trap mass spectrometry, with its capacity to perform multiple stages of fragmentation (MSn), is demonstrated as an effective method for the structural characterization of permethylated N-linked complex glycoprotein oligosaccharides. Complex glycan structural features, such as N-acetyllactosamine antenane, neuraminic acids, and nonreducing terminal GlcNAc monosaccharides, commonly suppress cross-ring and core saccharide cleavages in traditional MS/MS experiments. Using ion trap mass spectrometry, removal of these substituents permits determination of branching patterns and intersaccharide linkages by MS3 and MS4. Both sequence and linkage data are obtained for N-acetyllactosamine and sialyl-N-acetyllactosamine oligosaccharide antennae from biantennary glycans using MS3, and the location of a bisecting GlcNAc residue is also established after exposing the core pentasaccharide. Higher-order experiments further illustrate the potential of electrospray ionization-quadrupole ion trap mass spectrometry for carbohydrate analysis, as MS8 is used to produce significant and otherwise unobtainable branching information for an oligosaccharide from chicken ovalbumin. These studies constitute further evidence of the unique role that ion trap mass spectrometry can assume in the area of oligosaccharide analysis.
Weiskopf
A S
AS
Department of Chemistry, Northeastern University, Boston, Massachusetts 02115, USA.
Vouros
P
P
Harvey
D J
DJ
D000606:Amino Sugars / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D008970:Molecular Weight; D009005:Monosaccharides / Q000737:chemistry; D009438:Neuraminic Acids / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*
D000606:Amino Sugars; D006023:Glycoproteins; D009005:Monosaccharides; D009438:Neuraminic Acids; D009844:Oligosaccharides; C000458:N-acetyllactosamine