Structure determination of a sulfated N-glycans, candidate for a precursor of the selectin ligand in bovine lung.
To clarify the structure of non-sialic acid anionic residue on N-glycans in the mammalian tissues, we have isolated sialidase-resistant anionic residue on N-glycans from bovine lung. Analyses by partial acid hydrolysis and glycosidase digestions combined with a two-dimensional HPLC mapping method revealed that the major sialidase-resistant anionic N-glycan had a fucosylbianntenary core structure. The anionic residue was identified as a sulfate ester by methanolysis, anion-exchange chromatography, and mass spectrometry. The linkage position of the sulfate ester was the 6-position of the GlcNAc residue on the Manalpha1-6 branch. This conclusion was based on the results of glycosidase digestions followed by two-dimensional HPLC mapping. Furthermore, the disialylated form of this sulfated glycan was dominant, and no asialo form was detected. The structure of the major anionic N-glycan prepared from bovine lung and having a sulfate was proposed to be the pyridylamino derivative of Siaalpha2-3Galphalbeta1-4(HSO(3)-6)GlcNAcbeta1-2Manalpha1-6(Siaalpha2-3Galbeta1-4GlcNAcbeta1-2Manalpha1-3)Manbeta1-4GlcNAcbeta1-4(Fucalpha1-6)GlcNAc.
Murakami
Tomonori
T
Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka, 560-0043, Japan.
Natsuka
Shunji
S
Nakakita
Shin-Ichi
S
Hase
Sumihiro
S
D000818:Animals; D000838:Anions / Q000737:chemistry; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D008024:Ligands; D008168:Lung / Q000737:chemistry*; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D019087:Selectins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013463:Sulfuric Acid Esters / Q000737:chemistry*
D000838:Anions; D008024:Ligands; D011134:Polysaccharides; D019087:Selectins; D013463:Sulfuric Acid Esters; D009584:Nitrogen