Synthesis and testing of mechanism-based protein-profiling probes for retaining endoglycosidases

Abstract

New functional proteomics methods are required for targeting and identification of subsets of a proteome in an activity-based fashion. Glycosidases play critical roles in biology, yet a robust method for functional analysis of their activities and identities in biological proteomes is still lacking. An aryl 2-deoxy-2-fluoro xylo-bioside inactivator was conjugated through cleavable and non-cleavable linker arms to a biotin tag, thereby yielding two new active-site-directed reagents for activity-based profiling of retaining β-glycanases in complex proteomes. Crucially, these tagged reagents possess high specificity for their target enzymes with kinetic parameters similar to those of the untagg..