Activity-based protein profiling in vivo using a copper(i)-catalyzed azide-alkyne [3 + 2] cycloaddition.
Toward the goal of assigning function to the tens of thousands of protein products encoded by eukaryotic and prokaryotic genomes, the field of proteomics requires new technologies that can functionally characterize proteins within the dynamic environment of the cell, where these biomolecules are subject to myriad posttranslational modifications and the actions of endogenous activators and inhibitors. Here, we report an advanced strategy for activity-based protein profiling (ABPP) that addresses this important need. We show that several enzymes can be labeled in an activity-based manner both in vitro and in vivo by an azido-sulfonate ester probe and that these labeling events can be detected in whole proteomes by copper-catalyzed ligation with a rhodamine-alkyne reagent. This click chemistry-based strategy for ABPP represents a unique and versatile method for functional proteome analysis.
Speers
Anna E
AE
The Skaggs Institute for Chemical Biology, Departments of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Adam
Gregory C
GC
Cravatt
Benjamin F
BF
D000444:Aldehyde Dehydrogenase / Q000032:analysis*; D000080924:Aldehyde Dehydrogenase 1 Family; D000480:Alkynes / Q000737:chemistry*; D000818:Animals; D001386:Azides / Q000737:chemistry*; D001943:Breast Neoplasms / Q000201:enzymology; D019556:COS Cells; D002384:Catalysis; D002522:Chlorocebus aethiops; D003300:Copper / Q000737:chemistry*; D005982:Glutathione Transferase / Q000032:analysis*; D006801:Humans; D007527:Isoenzymes / Q000032:analysis*; D008099:Liver / Q000201:enzymology; D051379:Mice; D040901:Proteomics / Q000379:methods*; D050697:Retinal Dehydrogenase; D013329:Structure-Activity Relationship; D014162:Transfection; D014407:Tumor Cells, Cultured
D000480:Alkynes; D001386:Azides; D007527:Isoenzymes; D003300:Copper; D000080924:Aldehyde Dehydrogenase 1 Family; D000444:Aldehyde Dehydrogenase; C510223:ALDH1A1 protein, human; C515028:ALDH1A1 protein, mouse; D050697:Retinal Dehydrogenase; D005982:Glutathione Transferase