Structural studies on heparan sulphate from human lung fibroblasts. Characterization of oligosaccharides obtained by selective periodate oxidation of D-glucuronic acid residues followed by scission in alkali.
Structural studies on heparan sulphate from human lung fibroblasts. Characterization of oligosaccharides obtained by selective periodate oxidation of D-glucuronic acid residues followed by scission in alkali.
复制标题
人肺成纤维细胞硫酸乙酰肝素的结构研究。
DOI：
10.1042/bj1910103
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
L. Fransson
中科院分区：
文献类型：
--
作者：
I. Sjöberg;L. Fransson
1. (3)H-和（35）从人肺成纤维细胞单层中分离S-标记的硫酸乙酰肝素，并通过（a）脱氨基裂解和（B）高碘酸盐氧化/碱性消除进行降解。通过凝胶色谱和离子交换色谱分离片段。2.放射性聚糖的脱氨基裂解主要得到具有低含量的酯-硫酸酯和游离硫酸酯的二糖，这表明大部分（约10%）的多糖是由单糖组成的。80%）的重复单元由糖醛酸基-葡糖胺-N-硫酸盐组成。非硫酸化[葡萄糖醛酸基-N-乙酰葡糖胺]重复单元（3-4个连续单元）的嵌段占链的其余部分。3.通过选择性氧化与N-乙酰葡糖胺相关的葡糖醛酸残基，然后在碱中断裂，放射性聚糖被降解成一系列片段。含葡萄糖醛酸基-N-乙酰葡糖胺的嵌段区域产生化合物N-乙酰葡糖胺-R，其中R是氧化和降解的葡萄糖醛酸的残余物。高碘酸盐不敏感的糖醛酸残基在一般结构葡糖胺-（糖醛酸-葡糖胺）（n）-R的中间体中回收。4.通过脱氨基裂解和用高碘酸盐再氧化进一步降解这些葡萄糖醛酸，表明艾杜糖醛酸可能位于葡萄糖胺-N-硫酸盐-->艾杜糖醛酸--> N-乙酰葡萄糖胺的序列中。偶尔艾杜糖醛酸被硫酸化。含艾杜糖醛酸重复序列的嵌段可含有多达五个连续单元。还观察到艾杜糖醛酸和葡萄糖醛酸重复序列的交替排列。5. (3)H-和（35）来自成纤维细胞的连续提取物（培养基、EDTA、胰蛋白酶消化物、二硫苏糖醇提取物、细胞可溶性和细胞不溶性材料）的S-标记硫酸乙酰肝素在高碘酸盐氧化/碱消除和脱氨基裂解后提供了相似的曲线。
1. (3)H- and (35)S-labelled heparan sulphate was isolated from monolayers of human lung fibroblasts and subjected to degradations by (a) deaminative cleavage and (b) periodate oxidation/alkaline elimination. Fragments were resolved by gel- and ion-exchange-chromatography. 2. Deaminative cleavage of the radioactive glycan afforded mainly disaccharides with a low content of ester-sulphate and free sulphate, indicating that a large part (approx. 80%) of the repeating units consisted of uronosyl-glucosamine-N-sulphate. Blocks of non-sulphated [glucuronosyl-N-acetyl glucosamine] repeats (3-4 consecutive units) accounted for the remainder of the chains. 3. By selective oxidation of glucuronic acid residues associated with N-acetylglucosamine, followed by scission in alkali, the radioactive glycan was degraded into a series of fragments. The glucuronosyl-N-acetylglucosamine-containing block regions yielded a compound N-acetylglucosamine-R, where R is the remnant of an oxidized and degraded glucuronic acid. Periodate-insensitive uronic acid residues were recovered in saccharides of the general structure glucosamine-(uronic acid-glucosamine)(n)-R. 4. Further degradations of these saccharides via deaminative cleavage and re-oxidations with periodate revealed that iduronic acid may be located in sequences such as glucosamine-N-sulphate-->iduronic acid-->N-acetylglucosamine. Occasionally the iduronic acid was sulphated. Blocks of iduronic acid-containing repeats may contain up to five consecutive units. Alternating arrangements of iduronic acid- and glucuronic acid-containing repeats were also observed. 5. (3)H- and (35)S-labelled heparan sulphates from sequential extracts of fibroblasts (medium, EDTA, trypsin digest, dithiothreitol extract, cell-soluble and cell-insoluble material) afforded similar profiles after both periodate oxidation/alkaline elimination and deaminative cleavage.