Sojar H T, Bahl O P
Department of Biological Sciences, State University of New York at Buffalo 14260.
Arch Biochem Biophys. 1987 Nov 15;259(1):52-7. doi: 10.1016/0003-9861(87)90469-3.
A simple and rapid chemical method for the deglycosylation of glycoproteins has been developed. The method involves the incubation of protein with trifluoromethanesulfonic acid at 0 degrees C from 0.5 to 2 h followed by the neutralization of the acid with aqueous pyridine at -20 degrees C. The method has been applied effectively to fetuin, ovine submaxillary mucin, ovine lutropin, and human choriogonadotropin. In 1 h almost all of N- and O-linked carbohydrates from ovine lutropin and human choriogonadotropin, with the exception of the linkage N-acetylglucosamine or N-acetylgalactosamine, were removed. Similarly, in 1 h all N-linked carbohydrates, excepting again the linkage sugar, in fetuin were degraded. Longer reaction times up to 2 h completely removed the O-linked carbohydrate chains from fetuin and ovine submaxillary mucin. The deglycosylated hormones thus prepared retained their immunological and biological activities.
已开发出一种用于糖蛋白去糖基化的简单快速化学方法。该方法包括将蛋白质与三氟甲磺酸在0℃下孵育0.5至2小时,然后在-20℃下用吡啶水溶液中和酸。该方法已有效地应用于胎球蛋白、绵羊下颌粘液蛋白、绵羊促黄体激素和人绒毛膜促性腺激素。在1小时内,除连接的N-乙酰葡糖胺或N-乙酰半乳糖胺外,绵羊促黄体激素和人绒毛膜促性腺激素中几乎所有的N-和O-连接碳水化合物都被去除。同样,在1小时内,胎球蛋白中除连接糖外的所有N-连接碳水化合物都被降解。长达2小时的较长反应时间完全去除了胎球蛋白和绵羊下颌粘液蛋白中的O-连接碳水化合物链。由此制备的去糖基化激素保留了它们的免疫和生物活性。