The Asn‐linked carbohydrate chains of human Tamm‐Horsfall glycoprotein of one male

Abstract

Human Tamm‐Horsfall glycoprotein has been purified from the urine of one male. The Asn‐linked carbohydrate chains were enzymically released by peptide‐ N 4 ‐( N ‐acetyl‐β‐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‐NH 2 , 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 1 H‐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: