Application of negative ion MS/MS to the identification of N-glycans released from carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1).
Structures of N-glycans released from rat CEACAM1 expressed in human embryonic kidney cells were determined by MALDI and negative ion nanospray MS/MS techniques. The major carbohydrates were bi-, tri- and tetra-antennary complex glycans with and without sialic acid, fucose and bisecting GlcNAc residues. High-mannose glycans, predominantly Man(5)GlcNAc(2), were also found. The negative ion fragmentation technique easily identified the branching pattern of the triantennary glycans (mainly branched on the 6-antenna) and the presence of 'bisecting' GlcNAc residues (attached to the 4-position of the core mannose), features that are difficult to determine by traditional techniques. Sialic acids were in both alpha2-3 and alpha2-6 linkage as determined by MALDI-TOF MS following linkage-specific derivatization.
Harvey
David J
DJ
Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK. david.harvey@bioch.ox.ac.uk
Baruah
Kavitha
K
Scanlan
Christopher N
CN
D000818:Animals; D015703:Antigens, CD / Q000737:chemistry* / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000378:metabolism; D002460:Cell Line; D005643:Fucose / Q000737:chemistry; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D008358:Mannose / Q000737:chemistry; D015394:Molecular Structure; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D011134:Polysaccharides; D005643:Fucose; D019158:N-Acetylneuraminic Acid; D008358:Mannose