Development of bestatin-based activity-based probes for metallo-aminopeptidases.
A novel set of activity-based probes (ABPs) for functionally profiling metallo-aminopeptidases was synthesized based on the bestatin inhibitor scaffold, the first synthesis of bestatin analogues using solid-phase techniques. These ABPs were shown to label metallo-aminopeptidases, using both a biotin and a fluorophore reporter, in an activity-dependent manner. This probe class was also shown to be amenable to 'click' chemistry labeling for possible use in live cells. Finally, we demonstrate that the ABPs are able to label an aminopeptidase in a complex proteome. Thus, these bestatin-based probes should have wide utility to functionally profile aminopeptidases in many biological systems.
Harbut
Michael B
MB
University of Pennsylvania, Department of Pharmacology, 433 S. University Avenue, 304G Lynch Laboratories, Philadelphia, PA 19104-6018, USA.
Velmourougane
Geetha
G
Reiss
Gilana
G
Chandramohanadas
Rajesh
R
Greenbaum
Doron C
DC
D000626:Aminopeptidases / Q000378:metabolism*; D020134:Catalytic Domain; D005456:Fluorescent Dyes; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008958:Models, Molecular; D015394:Molecular Structure; D013329:Structure-Activity Relationship
D005456:Fluorescent Dyes; D000626:Aminopeptidases; D007930:Leucine; C012211:ubenimex