A chemical method for the deglycosylation of proteins.
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.
Sojar
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Department of Biological Sciences, State University of New York at Buffalo 14260.
Bahl
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D000596:Amino Acids / Q000032:analysis; D002241:Carbohydrates / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins; D006868:Hydrolysis; D008698:Mesylates
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D008698:Mesylates; C012077:trifluoromethanesulfonic acid