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

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• 作者：Bindu Nanduri^a ; Leslie A. Shack^a ; Aswathy N. Rai^a ; William B. Epperson^b ; Wes Baumgartner^b ; Ty B. Schmidt^c ; Mariola J. Edelmann^d ; ^{medelmann@ufl.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Deubiquitinases ; Focused-ultrasonication ; Activity-based profiling ; Chemical proteomics ; Tissue lysis
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：15 December 2016
• 年：2016
• 卷：515
• 期：Complete
• 页码：9-13
• 全文大小：813 K

文摘

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.