Isolation and structural characterization of sialic-acid-containing glycopeptides of the O-glycosidic type from the urine of two patients with an hereditary deficiency in alpha-N-acetylgalactosaminidase activity.
Glycopeptides have been isolated from the urine of two patients, aged 5 and 6, with a new lysosomal storage disease characterized by a deficiency in alpha-N-acetylgalactosaminidase activity. Isolation of these glycopeptides was achieved using gel filtration and ion-exchange chromatography. Structural determination was done using one- and two-dimensional 500 MHz 1H-NMR spectroscopy and FAB mass spectrometry of native and derivatized glycopeptides. The following structures were inferred as being present: Glycopeptide A (up to 140 mg/l urine) (1)-(3) Neu5Ac alpha 2-3Gal beta 1-3 (Neu5Ac alpha 2-6)GalNAc alpha 1-R A1: R = Ser A2: R = Thr A3: R = Thr-Pro Glycopeptide B (up to 80 mg/l urine) (4)-(6) Neu5Ac alpha 2-3Gal beta 1-4GlcNAc beta 1-6 (Neu5Ac alpha 2-3-Gal beta 1-3) GalNAc alpha 1-R B1: R = Ser B2:R = Thr B3: R = Thr-Pro
Institut für Physiologische Chemie, Universität Bonn.
H U
HU
Linden
R A
RA
Klein
H
H
Egge
J
J
Peter-Katalinic
J
J
Dabrowski
D
D
Schindler
D000596:Amino Acids / Q000032:analysis; D001894:Borohydrides; D002239:Carbohydrate Metabolism, Inborn Errors / Q000378:metabolism; D002850:Chromatography, Gel; D006020:Glycopeptides / Q000652:urine*; D006027:Glycosides / Q000652:urine*; D006596:Hexosaminidases / Q000172:deficiency*; D006801:Humans; D006868:Hydrolysis; D008247:Lysosomes / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008745:Methylation; D008968:Molecular Conformation; D011134:Polysaccharides / Q000378:metabolism; D012794:Sialic Acids / Q000652:urine*; D048809:alpha-N-Acetylgalactosaminidase
D000596:Amino Acids; D001894:Borohydrides; D006020:Glycopeptides; D006027:Glycosides; D011134:Polysaccharides; D012794:Sialic Acids; D006596:Hexosaminidases; C482722:NAGA protein, human; D048809:alpha-N-Acetylgalactosaminidase