Comparative analysis of click chemistry mediated activity-based protein profiling in cell lysates

Activity-based protein profiling uses chemical probes that covalently attach to active enzyme targets. Probes with conventional tags have disadvantages, such as limited cell permeability or steric hindrance around the reactive group. A tandem labeling strategy with click chemistry is now widely used...

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Published inMolecules (Basel, Switzerland) Vol. 18; no. 10; pp. 12599 - 12608
Main Authors Yang, Yinliang, Yang, Xiaomeng, Verhelst, Steven H L
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 11.10.2013
MDPI
Subjects
activity-based probes
Alkynes - chemistry
Animals
Azide
Azides - chemistry
Biocatalysis
cathepsins
Cathepsins - chemistry
Cell Extracts - chemistry
Cell Fractionation
Cell Line
Chemical bonds
Chemistry
Click Chemistry
Communication
Comparative analysis
Copper Sulfate - chemistry
Efficiency
Enzymes
Labeling
Ligands
Mice
Permeability
proteases
protein modification
Proteins
Staining and Labeling
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Summary:Activity-based protein profiling uses chemical probes that covalently attach to active enzyme targets. Probes with conventional tags have disadvantages, such as limited cell permeability or steric hindrance around the reactive group. A tandem labeling strategy with click chemistry is now widely used to study enzyme targets in situ and in vivo. Herein, the probes are reacted in live cells, whereas the ensuing detection by click chemistry takes place in cell lysates. We here make a comparison of the efficiency of the activity-based tandem labeling strategy by using Cu(I)-catalyzed and strain-promoted click chemistry, different ligands and different lysis conditions.
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ISSN:1420-3049
1420-3049
DOI:10.3390/molecules181012599