Construction of linear GlcNAc beta 1-6Gal beta 1-OR type oligosaccharides by partial cleavage of GlcNAc beta 1-3(GlcNAc beta 1-6)Gal beta 1-OR sequences with jack bean beta-N-acetylhexosaminidase.
Radiolabelled GlcNAc beta 1-3(GlcNAc beta 1-6)Gal (1), GlcNAc beta 1-3)GlcNAc beta 1-6)Gal beta 1-OCH3 (4), GlcNAc beta 1-3(GlcNAc beta 1-6)Gal beta 1-4Glc (7), and GlcNAc beta 1-3(GlcNAc beta 1-6)Gal beta 1-4GlcNAc (10) were cleaved partially with jack bean beta-N-acetylhexosaminidase (EC 3.2.1.30), and the digests were analysed chromatographically. All four oligosaccharides were hydrolysed faster at the (1-6) branch, than at the (1-3) branch, but a high branch specificity was observed only with the glycan 4. The saccharides 1 and 7 resembled each other in the kinetics of the enzyme-catalysed release of their two non-reducing N-acetylglucosamine units, but the glycan 10 was rather different. The partial digestions made it possible to obtain radiolabelled GlcNAc beta 1-6Gal, GlcNAc beta 1-6Gal beta 1-OCH3, GlcNAc beta 1-6Gal beta 1-4Glc, and, in particular, GlcNAc beta 1-6Gal beta 1-4GlcNAc.
Renkonen
O
O
Institute of Biotechnology, University of Helsinki, Finland.
Niemelä
R
R
Leppänen
A
A
Maaheimo
H
H
Seppo
A
A
Penttilä
L
L
Vilkman
A
A
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002854:Chromatography, Paper; D006868:Hydrolysis; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism*
D009844:Oligosaccharides; D001619:beta-N-Acetylhexosaminidases