Use of focused ultrasonication in activity-based profiling of deubiquitinating enzymes in tissue

作者：Nanduri Bindu; Shack Leslie A; Rai Aswathy N; Epperson William B; Baumgartner Wes; Schmidt Ty B; Edelmann Mariola J^*

来源：Analytical Biochemistry, 2016, 515: 9-13.

DOI：10.1016/j.ab.2016.09.016

• Deubiquitinases
• Focused-ultrasonication
• Activity-based profiling
• Chemical proteomics
• Tissue lysis

To develop a reproducible tissue lysis method that retains enzyme function for activity-based protein profiling, we compared four different methods to obtain protein extracts from bovine lung tissue: focused ultrasonication, standard sonication, mortar & pestle method, and homogenization combined with standard sonication. Focused ultrasonication and mortar & pestle methods were sufficiently effective for activity-based profiling of deubiquitinases in tissue, and focused ultrasonication also had the fastest processing time. We used focused-ultrasonicator for subsequent activity-based proteomic analysis of deubiquitinases to test the compatibility of this method in sample preparation for activity-based chemical proteomics.

• 出版日期2016-12-15