학술논문
Use of focused ultrasonication in activity-based profiling of deubiquitinating enzymes in tissue.
Document Type
Article
Author
Source
Subject
*ULTRASONICS
*ENZYMES
*PROTEINS
*PROTEOMICS
*ASYMPTOTIC homogenization
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Language
ISSN
0003-2697
Abstract
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. [ABSTRACT FROM AUTHOR]