Abstract
Functional glycomics has been impeded by the lack of inexpensive enzymatic and mild chemical methods to acquire natural glycans in significant amounts. In this study, we have developed a new strategy we term "threshing and trimming" (TaT) to quickly obtain N-glycans from glycoproteins and animal tissues. TaT employs low-cost Pronase to degrade peptides and N-bromosuccinimide (NBS) to effect oxidative decarboxylation under very mild reaction conditions to generate homogeneous aglycon moieties as nitriles or aldehydes. These aglycons can be readily conjugated with fluorescent tags for profiling and functional study. TaT is an affordable alternative to expensive specialty enzymes and strong chemical treatment and unpleasant reagents, and should further drive the functional glycomics of N-glycans.