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小牛Tamm-Horsfall糖蛋白的制备及化学特性分析

Preparation and chemical characterisation of calf Tamm-Horsfall glycoprotein.

作者信息

van Dijk W, Lasthuis A M, Ferwerda W

出版信息

Biochim Biophys Acta. 1979 Apr 18;584(1):121-8. doi: 10.1016/0304-4165(79)90242-3.

Abstract

Tamm-Horsfall glycoprotein preparations were obtained from calf urine by 1.0 M NaCl precipitation followed by 4 M urea/Sepharose 4B chromatography. By using 0.1% sodium dodecyl sulfate polyacrylamide gel electrophoresis a molecular weight of 86 500 +/- 4500 (n = 12) was calculated for the glycoprotein. Amino acid and carbohydrate analyses were performed, the carbohydrate composition being (in residues per 100 amino acid residues in the glycoprotein): fucose, 0.90; galactose, 4.82; mannose, 4.63;N-acetylglucosamine, 7.36; N-acetylgalactosamine, 1.38; sialic acid, 2.93. Under conditions of mild acid hydrolysis (0.05 M H2SO4, 80 degrees C, 1 h) the calf Tamm-Horsfall glycoprotein preparations were degraded partially into two lower molecular weight fragments (approximate Mr 66 000 and 51 000), as shown by polyacrylamide gel electrophoresis, both fragments being periodic acid-Schiff reagent positive.

摘要

通过用1.0M氯化钠沉淀,随后进行4M尿素/琼脂糖4B柱色谱法,从小牛尿液中获得了Tamm-Horsfall糖蛋白制剂。通过使用0.1%十二烷基硫酸钠聚丙烯酰胺凝胶电泳,计算出该糖蛋白的分子量为86500±4500(n=12)。进行了氨基酸和碳水化合物分析,碳水化合物组成(以糖蛋白中每100个氨基酸残基中的残基计)为:岩藻糖,0.90;半乳糖,4.82;甘露糖,4.63;N-乙酰葡糖胺,7.36;N-乙酰半乳糖胺,1.38;唾液酸,2.93。在温和酸水解条件下(0.05M硫酸,80℃,1小时),小牛Tamm-Horsfall糖蛋白制剂部分降解为两个较低分子量的片段(约Mr 66000和51000),如聚丙烯酰胺凝胶电泳所示,两个片段均对高碘酸-席夫试剂呈阳性。

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