Use of focused ultrasonication in activity-based profiling of deubiquitinating enzymes in tissue.
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.
College of Veterinary Medicine, Department of Basic Sciences, Mississippi State University, MS, USA.
Bindu
B
Nanduri
College of Veterinary Medicine, Department of Basic Sciences, Mississippi State University, MS, USA.
Leslie A
LA
Shack
College of Veterinary Medicine, Department of Basic Sciences, Mississippi State University, MS, USA.
Aswathy N
AN
Rai
College of Veterinary Medicine, Department of Pathobiology and Population Medicine, Mississippi State University, MS, USA.
William B
WB
Epperson
College of Veterinary Medicine, Department of Pathobiology and Population Medicine, Mississippi State University, MS, USA.
Wes
W
Baumgartner
Animal Science Department, Institute of Agriculture and Natural Resources, University of Nebraska - Lincoln, USA.
Ty B
TB
Schmidt
Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, USA. Electronic address: medelmann@ufl.edu.
Mariola J
MJ
Edelmann
D000818:Animals; D002417:Cattle; D000072017:Deubiquitinating Enzymes / Q000737:chemistry*; D008168:Lung / Q000201:enzymology*; D040901:Proteomics / Q000379:methods*; D000069453:Ultrasonic Waves
D000072017:Deubiquitinating Enzymes
Activity-based profiling; Chemical proteomics; Deubiquitinases; Focused-ultrasonication; Tissue lysis
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