The Asn-linked carbohydrate chains of human Tamm-Horsfall glycoprotein of one male. Novel sulfated and novel N-acetylgalactosamine-containing N-linked carbohydrate chains.
Human Tamm-Horsfall glycoprotein has been purified from the urine of one male. The Asn-linked carbohydrate chains were enzymically released by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F, and separated from the remaining protein by gel-permeation chromatography on Bio-Gel P-100. Fractionation of the intact (sulfated) sialylated carbohydrate chains was achieved by a combination of three liquid-chromatographic techniques, namely, anion-exchange FPLC on Q-Sepharose, amine-adsorption HPLC on Lichrospher-NH2, and high-pH anion-exchange chromatography on CarboPac PA1. In total, more than 150 carbohydrate-containing fractions were obtained, some of which still contained mixtures of oligosaccharides. The primary structure of 30 N-glycans, including 10 novel oligosaccharides, were determined by one- and two-dimensional 1H-NMR spectroscopy at 500 MHz or 600 MHz. The types of compounds identified range from non-fucosylated, monosialylated, diantennary to fucosylated, tetrasialylated, tetraantennary carbohydrate chains, possessing the following terminal structural elements: [formula: see text]
Hård
K
K
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
Van Zadelhoff
G
G
Moonen
P
P
Kamerling
J P
JP
Vliegenthart
F G
FG
D000116:Acetylgalactosamine / Q000737:chemistry*; D001216:Asparagine / Q000737:chemistry*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008969:Molecular Sequence Data; D009088:Mucoproteins / Q000737:chemistry*; D013431:Sulfates / Q000737:chemistry*; D058949:Uromodulin
D002241:Carbohydrates; D009088:Mucoproteins; D013431:Sulfates; C548612:UMOD protein, human; D058949:Uromodulin; D001216:Asparagine; D000116:Acetylgalactosamine