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Activity-based protein profiling in vivo using a copper(i)-catalyzed azide-alkyne [3 + 2] cycloaddition
Anna E Speers1, Gregory C Adam, Benjamin F Cravatt
1The Skaggs Institute for Chemical Biology, Departments of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|April 17, 2003
Summary
This study introduces a new activity-based protein profiling (ABPP) method using click chemistry to label and detect active enzymes in cells. This advanced technique enables functional proteome analysis for better protein function assignment.
Area of Science:
- Proteomics
- Biochemistry
- Molecular Biology
Background:
- Assigning functions to proteins encoded by genomes is a key challenge in proteomics.
- Characterizing protein function within the cell requires technologies that account for posttranslational modifications and regulatory factors.
Purpose of the Study:
- To develop an advanced strategy for activity-based protein profiling (ABPP) to functionally characterize proteins in vivo.
- To enable the detection of active enzymes within the complex cellular environment.
Main Methods:
- Utilized an azido-sulfonate ester probe for activity-based labeling of enzymes in vitro and in vivo.
- Employed copper-catalyzed ligation with a rhodamine-alkyne reagent for detection of labeled proteins.
- Applied click chemistry for functional proteome analysis.
Main Results:
- Demonstrated successful activity-based labeling of several enzymes using the azido-sulfonate ester probe.
- Showcased the detection of these labeling events in whole proteomes.
- Validated the efficacy of the click chemistry-based ABPP strategy.
Conclusions:
- The developed click chemistry-based ABPP strategy is a versatile method for functional proteome analysis.
- This approach facilitates the assignment of functions to a large number of protein products.
- Provides a novel tool for studying protein activity in dynamic cellular environments.
