title: Comparative analysis of click chemistry mediated activity-based protein profiling in cell lysates.
abstract: 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.
authors.affiliation: Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany. verhelst@wzw.tum.de.
authors.forename: Yinliang
authors.initials: Y
authors.lastname: Yang
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Yang
authors.forename: Steven H L
authors.initials: SH
authors.lastname: Verhelst
mesh_terms: D000480:Alkynes / Q000737:chemistry; D000818:Animals; D001386:Azides / Q000737:chemistry; D055162:Biocatalysis; D002403:Cathepsins / Q000737:chemistry*; D002457:Cell Extracts / Q000737:chemistry*; D002458:Cell Fractionation; D002460:Cell Line; D057930:Click Chemistry; D019327:Copper Sulfate / Q000737:chemistry; D008024:Ligands; D051379:Mice; D013194:Staining and Labeling
chemical_list: D000480:Alkynes; D001386:Azides; D002457:Cell Extracts; D008024:Ligands; D002403:Cathepsins; D019327:Copper Sulfate