Goso Y, Hotta K
Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara, Japan.
Biochem J. 1989 Dec 15;264(3):805-12. doi: 10.1042/bj2640805.
Radiolabelled mucus glycoprotein was obtained from tissue and a culture medium each of the corpus and antrum of rat stomach incubated with [35S]sulphate in vitro. Gel-filtration analysis of oligosaccharides liberated by alkaline-borohydride treatment from glycoproteins indicated that 35S-labelled oligosaccharides from the corpus vary considerably with respect to chain length whereas those from antral mucus glycoprotein are composed of small oligosaccharides. Examination of the reduced radiolabelled products obtained by HNO2 cleavage of the hydrazine-treated oligosaccharides indicated sulphate esters of N-acetylglucosamine to be present at three locations on a carbohydrate unit: [35S]sulphated monosaccharide (2,5-anhydromannitol 6-sulphate), [35S]sulphated disaccharide [galactosyl(beta 1-4)-2,5-anhydromannitol 6-sulphate] and [35S]sulphated trisaccharide [fucosyl(alpha 1-2)-galactosyl(beta 1-4)-2,5-anhydromannitol 6-sulphate]. Sulphated disaccharide and trisaccharide, possibly originating from the N-acetyl-lactosamine and fucosyl-N-acetyl-lactosamine sequences respectively, were detected in the corpus, especially as large oligosaccharides, but were present in the antrum in only very small amounts. The sulphated monosaccharide, however, most probably originating from 6-sulphated N-acetylglucosamine residues at non-reducing termini, was present in all oligosaccharide fractions in both the corpus and antrum.
将大鼠胃体和胃窦的组织及培养基分别在体外与[35S]硫酸盐一起孵育,获得放射性标记的黏液糖蛋白。对经碱性硼氢化物处理从糖蛋白释放的寡糖进行凝胶过滤分析表明,胃体中35S标记的寡糖在链长方面差异很大,而胃窦黏液糖蛋白中的寡糖由小寡糖组成。对经肼处理的寡糖经亚硝酸裂解得到的还原放射性标记产物进行检查,结果表明在碳水化合物单元的三个位置存在N-乙酰葡糖胺的硫酸酯:[35S]硫酸化单糖(2,5-脱水甘露糖醇6-硫酸盐)、[35S]硫酸化二糖[半乳糖基(β1-4)-2,5-脱水甘露糖醇6-硫酸盐]和[35S]硫酸化三糖[岩藻糖基(α1-2)-半乳糖基(β1-4)-2,5-脱水甘露糖醇6-硫酸盐]。硫酸化二糖和三糖可能分别源自N-乙酰乳糖胺和岩藻糖基-N-乙酰乳糖胺序列,在胃体中被检测到,尤其是作为大寡糖,但在胃窦中仅以极少量存在。然而,硫酸化单糖很可能源自非还原末端的6-硫酸化N-乙酰葡糖胺残基,在胃体和胃窦的所有寡糖组分中均有存在。