Global Analysis of Cellular Proteolysis by Selective Enzymatic Labeling of Protein N-Termini

摘要: CHAPTER THIRTEEN Global Analysis of Cellular Proteolysis by Selective Enzymatic Labeling Protein N-Termini Arun P. Wiita * ,†,1 , Julia E. Seaman ,1 James A. Wells ,{,2 Department Pharmaceutical Chemistry, University California, San Francisco, USA Laboratory Medicine, and Molecular Pharmacology, These authors contributed equally to this work. Corresponding author: e-mail address: jim.wells@ucsf.edu Contents 1. Introduction 1.1 Importance proteolysis 1.2 Approaches substrate cleavages identification 2. Basic Features Subtiligase Method 2.1 2.2 Types samples peptide tags 2.3 Sample setup introduction 3. Subtiligase-Based 3.1 Overview method 3.2 Specialized reagents for subtiligase labeling enrichment 4. Experimental Applications N-Terminomics 4.1 Application cell culture systems undergoing apoptosis (Forward Discovery experiments) 4.2 Identifying specific substrates recombinant proteases in lysates (Reverse 4.3 Quantification kinetics 4.4 human plasma serum 4.5 N-terminal any biological sample 5. Limitations the 6. Summary Findings from 7. Future Directions Acknowledgments References Methods Enzymology, Volume 544 ISSN 0076-6879 http://dx.doi.org/10.1016/B978-0-12-417158-9.00013-3 2014 Elsevier Inc. All rights reserved.