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从大鼠白血病(RBL-1)嗜碱性粒细胞中分离和鉴定硫酸化粘多糖

Isolation and characterization of sulphated mucopolysaccharides from rat leukaemic (RBL-1) basophils.

作者信息

Metcalfe D D, Wasserman S I, Austen K F

出版信息

Biochem J. 1980 Feb 1;185(2):367-72. doi: 10.1042/bj1850367.

Abstract

Proteoglycans of 300 000 mol.wt. were isolated from dispersed rat basophil tumour cells after labelling of the sulphated mucopolysaccharides with 35S in vitro:90% of the 35S-labelled mucopolysaccharides were extracted at high salt concentration. Alkali degradation of the 35S-labelled proteoglycans yielded glycosaminoglycan chains of 40 000 mol.wt. The composition of the salt-extracted 35S-labelled mucopolysaccharides, as defined by parallel or sequential degradation with chondroitinase AC, chondroitinase ABC and heparinase and resolution of the disaccharide-digestion products obtained with chondroitinase AC, was 48--61% chondroitin 4-sulphate, 20--30% dermatan sulphate, 10--15% heparin and 7--9% chondroitin 6-sulphate. Most of the salt-extracted 35S-labelled mucopolysaccharides were highly charged, with heparin and chondroitin 6-sulphate being relatively uniform in this regard, whereas chondroitin 4-sulphate and dematan sulphate exhibited a range of charge characteristics. The diversity of sulphated mucopolysaccharides present in the rat leukaemic basophil is in contrast with the predominance of heparin in the rat mast cell.

摘要

在体外以35S标记硫酸化粘多糖后,从分散的大鼠嗜碱性粒细胞肿瘤细胞中分离出分子量为300000的蛋白聚糖:90%的35S标记粘多糖在高盐浓度下被提取出来。对35S标记的蛋白聚糖进行碱降解,得到分子量为40000的糖胺聚糖链。通过用软骨素酶AC、软骨素酶ABC和肝素酶进行平行或顺序降解,并对软骨素酶AC得到的二糖消化产物进行分离,确定盐提取的35S标记粘多糖的组成如下:48% - 61%为硫酸软骨素4,20% - 30%为硫酸皮肤素,10% - 15%为肝素,7% - 9%为硫酸软骨素6。大部分盐提取的35S标记粘多糖带电量很高,在这方面肝素和硫酸软骨素6相对较为一致,而硫酸软骨素4和硫酸皮肤素则表现出一系列电荷特征。大鼠白血病嗜碱性粒细胞中存在的硫酸化粘多糖的多样性与大鼠肥大细胞中肝素占主导地位形成对比。

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