| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1183628 | EuPA Open Proteomics | 2014 | 7 Pages |
•We describe a method for in cell activity based protein profiling.•The method is applied to E. coli.•We compare the efficiency and patterns of labeling in cell vs in lysate.•Mainly qualitative differences are noted.•We perform mass spectrometry to identify the labeled proteins.
A fluorophosphonate based alkyne activity probe was used for the selective labeling of active serine hydrolases in intact Escherichia coli cells. A biotin-azide tag was subsequently attached to the alkyne functionality of the probe with copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. Comparison of proteins from in-cell and lysate labeled preparations suggested qualitatively similar patterns of reactivity in both preparations. Approximately 68%, 30 of the total 44 serine hydrolases detectable in E. coli were labeled with the probe indicating significant coverage with a single probe. The methods described here offer a useful tool for profiling and monitoring serine hydrolase activity in situ.
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