DEGLYCOSYLATION OF GLYCOPROTEINS BY TRIFLUOROMETHANESULFONIC ACID
DEGLYCOSYLATION OF GLYCOPROTEINS BY TRIFLUOROMETHANESULFONIC ACID
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DOI：
10.1016/0003-2697(81)90168-8
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发表时间：
1981-01-01
影响因子：
2.9
通讯作者：
WEBER, P
中科院分区：
文献类型：
--
作者：
EDGE, ASB;FALTYNEK, CR;WEBER, P
描述了一种糖蛋白化学去糖基化的新方法。在0℃下用三氟甲磺酸处理胎球蛋白。或25度。 C导致外周糖快速裂解，丝氨酸和苏氨酸连接的N-乙酰半乳糖胺缓慢损失，并保留N-糖苷连接的N-乙酰葡糖胺。氨基酸分析和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳表明，0℃反应后肽主链保持完整。 C 2.5 小时或在 25°C 下。 C 1小时。 0℃下处理绵羊黄体生成素和人M、N活性红细胞唾液酸糖蛋白。 C 3 小时给出相似的结果。就绵羊黄体生成激素而言，根据受体结合测定的测量，回收的去糖基化激素的活性约为起始材料的 2%。在牛鼻中隔蛋白多糖的情况下，去糖基化的核心蛋白充当UDP-D-木糖：核心蛋白β-O-木糖基转移酶的极好的受体。该试剂的毒性远低于 HF，并且不需要特殊设备进行处理。该程序不仅可用于生成去糖基化蛋白质，而且还可用于评估糖蛋白中天冬酰胺连接的糖链的数量。
A new method for the chemical deglycosylation of glycoproteins is described. Treatment of fetuin with trifluoromethanesulfonic acid at 0.degree. or 25.degree. C results in rapid cleavage of peripheral sugars, slow loss of serine- and threonine-linked N-acetylgalactosamine and retention of N-glycosidically linked N-acetylglucosamine. Amino acid analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveal that the peptide backbone is left intact after reaction at 0.degree. C for 2.5 h or at 25.degree. C for 1 h. Treatment of ovine luteinizing hormone and human M,N-active erythrocyte sialoglycoproteins at 0.degree. C for 3 h gives similar results. In the case of ovine luteinizing hormone, the recovered deglycosylated hormone has approximately 2% of the starting material''s activity as measured in a receptor binding assay. In the case of bovine nasal septum proteoglycan, the deglycosylated core protein acts as an excellent acceptor for UDP-D-xylose:core protein .beta.-O-xylosyltransferase. The reagent is far less toxic than HF and does not require special equipment for handling. The procedure should prove useful not only for generation of deglycosylated proteins, but also for assessment of the number of asparagine-linked saccharide chains in a glycoprotein.