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大鼠颊部和腭部小唾液腺在体内和体外合成的硫酸化糖蛋白的分离、化学及生物学特性研究

Isolation, chemical and biological characterization of sulphated glycoproteins synthesized by rat buccal and palatal minor salivary glands in vivo and in vitro.

作者信息

Green D R, Embery G

机构信息

Department of Dental Sciences, University of Liverpool, England, U.K.

出版信息

Arch Oral Biol. 1987;32(6):391-9. doi: 10.1016/0003-9969(87)90073-2.

Abstract

35S-labelled sulphated glycoproteins (SGP) were isolated from these glands after the incorporation of radiosulphate in vivo and in vitro by fractionation of tissue and medium extracts on Sepharose 4B and partial purification by DEAE-Sephacel anion exchange chromatography. Fractions were assessed for purity by SDS-PAGE and by cellulose-acetate electrophoresis. Molecular weights ranged from 34,000 to 5 X 10(6). It was notable that the molecular size of SGP from the in vitro media was generally lower than from the corresponding tissue fractions, particularly for the palatal samples. The fractions were heterogeneous and contained no sulphated glycosaminoglycans; they had high levels of aspartic acid, glutamic acid, threonine and serine, but there was no major difference in amino-acid composition between them. Carbohydrate analysis indicated typical components associated with sulphated glycoproteins, including fucose, galactose, glucose, mannose, N-acetylgalactosamine, N-acetylglucosamine and N-acetylneuraminic acid. Protein:carbohydrate ratios ranged from 0.1:1.0-3.5:1.0 and ester sulphate from 0.8 to 16.2 per cent. All fractions exhibited blood-group A reactivity and aggregated Streptococcus sanguis NCTC 7864; several fractions interacted similarly with Streptococcus mutans OMZ61.

摘要

通过在体内和体外将放射性硫酸盐掺入后,从这些腺体中分离出35S标记的硫酸化糖蛋白(SGP)。具体方法是对组织和培养基提取物在琼脂糖4B上进行分级分离,并通过DEAE - 琼脂糖凝胶阴离子交换色谱进行部分纯化。通过SDS - PAGE和醋酸纤维素电泳评估各组分的纯度。分子量范围为34,000至5×10⁶。值得注意的是,体外培养基中SGP的分子大小通常低于相应的组织组分,尤其是腭部样本。这些组分是异质的,不含硫酸化糖胺聚糖;它们含有高水平的天冬氨酸、谷氨酸、苏氨酸和丝氨酸,但它们之间的氨基酸组成没有重大差异。碳水化合物分析表明存在与硫酸化糖蛋白相关的典型成分,包括岩藻糖、半乳糖、葡萄糖、甘露糖、N - 乙酰半乳糖胺、N - 乙酰葡糖胺和N - 乙酰神经氨酸。蛋白质与碳水化合物的比例范围为0.1:1.0至3.5:1.0,酯硫酸盐含量为0.8%至16.2%。所有组分均表现出血型A反应性并能凝集血链球菌NCTC 7864;几个组分与变形链球菌OMZ61有类似的相互作用。

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