THE ASN-LINKED CARBOHYDRATE CHAINS OF HUMAN TAMM-HORSFALL GLYCOPROTEIN OF ONE MALE - NOVEL SULFATED AND NOVEL N-ACETYLGALACTOSAMINE-CONTAINING N-LINKED CARBOHYDRATE CHAINS
THE ASN-LINKED CARBOHYDRATE CHAINS OF HUMAN TAMM-HORSFALL GLYCOPROTEIN OF ONE MALE - NOVEL SULFATED AND NOVEL N-ACETYLGALACTOSAMINE-CONTAINING N-LINKED CARBOHYDRATE CHAINS
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DOI：
10.1111/j.1432-1033.1992.tb17362.x
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发表时间：
1992-11-01
期刊：
影响因子：
--
通讯作者：
VLIEGENTHART, JFG
中科院分区：
文献类型：
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作者：
HARD, K;VANZADELHOFF, G;VLIEGENTHART, JFG
人Tamm-Horsfall糖蛋白已从一名男性的尿液中纯化。天冬酰胺连接的碳水化合物链通过肽-N4-（N-乙酰基-β-葡糖胺基）天冬酰胺酰胺酶F酶促释放，并通过凝胶渗透色谱在Bio-Gel P-100上与剩余的蛋白质分离。通过组合三种液相色谱技术，即Q-Sepharose上的阴离子交换FPLC、Lichrospher-NH 2上的胺吸附HPLC和CarboPac PA 1上的高pH阴离子交换色谱，实现了完整（硫酸化）唾液酸化碳水化合物链的分馏。总共获得了超过150个含碳水化合物的部分，其中一些仍然含有低聚糖的混合物。通过一维和二维H-1-NMR光谱在500 MHz或600 MHz下测定了30个N-聚糖（包括10个新寡糖）的一级结构。所鉴定的化合物的类型范围从非岩藻糖基化、单唾液酸化、二触角到岩藻糖基化、四唾液酸化、四触角碳水化合物链，具有以下末端结构元素：[图形]最大的含GalNAc的化合物具有以下结构：（~）[图形]
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 H-1-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:[GRAPHICS]The largest GalNAc-containing compound has the following structure:(~)[GRAPHICS]