Ion-spray mass spectrometry for identification of the nonreducing terminal sugar of glycosaminoglycan.
Various oligosaccharides from hyaluronic acid, which have glucuronic acid or N- acetylglucosamine at the nonreducing terminal, were prepared by digestion with a combination of testicular hyaluronidase and beta-glucuronidase. These oligo saccharides were analyzed by negative-mode ion-spray mass spectrometry (MS) with an atmospheric pressure ion source. Introduction of collisionally activated dissociation tandem mass spectrometry (CAD-MS/MS) produced ions derived from cleavage of the glycosidic bonds, allowing the structure to be analyzed. The CAD-MS/MS spectrum showed an intense and characteristic fragment ion at m/z 193 for oligosaccharides having glucuronic acid at the nonreducing terminal. On the other hand, this ion was not observed in the spectra of oligosaccharides having N- acetylglucosamine at the nonreducing terminal. Therefore, the fragmentation pattern revealed by CAD-MS/MS provides useful information for distinguishing glucuronic acid and N- acetylglucosamine at the nonreducing terminal of oligosaccharides derived from hyaluronic acid and other glycosaminoglycans. This ion-spray CAD-MS/MS technique was also applied successfully to the characterization of glycosaminoglycans reconstructed by glycotechnology.
Takagaki
K
K
Department of Biochemistry, Hirosaki University School of Medicine, 5 Zaifu-cho, Hirosaki 036-8256, Japan.
Munakata
H
H
Nakamura
W
W
Matsuya
H
H
Majima
M
M
Endo
M
M
D000818:Animals; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis; D002417:Cattle; D004926:Escherichia coli / Q000201:enzymology; D005966:Glucuronidase / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry* / Q000302:isolation & purification; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods; D013737:Testis / Q000201:enzymology; D014470:Umbilical Cord / Q000737:chemistry
D002241:Carbohydrates; D006025:Glycosaminoglycans; D009844:Oligosaccharides; D006820:Hyaluronic Acid; D005966:Glucuronidase; D006821:Hyaluronoglucosaminidase