Comparative Analysis of Click Chemistry Mediated Activity-Based Protein Profiling in Cell Lysates

Molecules

Yinliang Yang¹  Xiaomeng Yang¹ 

[1] Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising, Germany;

关键词: activity-based probes;    cathepsins;    click chemistry;    proteases;    protein modification;   

DOI  :  10.3390/molecules181012599

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland.