Chemical Deglycosylation Can Induce Methylation, Succinimide Formation, and Isomerization

Abstract

Interpretation of deglycosylation studies relies heavily on the absence of modifications to the polypeptide chain. We have found that by using a common chemical deglycosylation technique, one can effect at least three changes in a peptide's structure: methylation, isomerization, and ring formation. It was determined that the conditions of chemical deglycosylation introduce a +14 Da shift in the masses of our model peptides, RKDVY, RKEVY, and horseradish peroxidase. This shift is localized to acidic functional groups and is interpreted as methylation of the free carboxylates in our models. An additional shift in mass of -18 Da is found in the model peptide RKDVY consistent with the loss of water associated with succinimide ring formation in this peptide. Chemical treatment induced isomerization of aspartyl residues to isoaspartyl residues in another model peptide, tetragastrin. These results indicate that one should use caution when interpreting the results of chemical deglycosylation experiments.