Ion-spray mass spectrometry for identification of the nonreducing terminal sugar of glycosaminoglycan

• Published in: Glycobiology (Oxford) Vol. 8; no. 7; pp. 719 - 724
• Main Authors: Takagaki, Keiichi, Munakata, Hidekazu, Nakamura, Wataru, Matsuya, Hideki, Majima, Mitsuo, Endo, Masahiko
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 01.07.1998
• Subjects: Animals; Carbohydrate Sequence; Carbohydrates - analysis; Cattle; Escherichia coli - enzymology; Glucuronidase - metabolism; glycosaminoglycan; Glycosaminoglycans - chemistry; glycotechnology; Humans; hyaluronic acid; Hyaluronic Acid - chemistry; Hyaluronic Acid - isolation & purification; Hyaluronoglucosaminidase - metabolism; ion-spray mass spectrometry; Male; Mass Spectrometry - methods; Molecular Sequence Data; nonreducing terminal sugar; Oligosaccharides - chemistry; Oligosaccharides - isolation & purification; Spectrometry, Mass, Secondary Ion - methods; Testis - enzymology; Umbilical Cord - chemistry
• ISSN: 0959-6658; 1460-2423
• DOI: 10.1093/glycob/8.7.719

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 β-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.