Characterization of the prime and non-prime active site specificities of proteases by proteome-derived peptide libraries and tandem mass spectrometry
Authors: Oliver Schilling, Pitter F. Huesgen, Olivier Barré, Ulrich auf dem Keller, Christopher M. Overall. Lab of Christopher Overall, Centre for Blood Research, Department of Oral Biological & Medical Sciences
Published Nature Protocols 6: 111-120 (2011). doi:10.1038/nprot.2010.178
Abstract: Knowledge of the cleavage site specificity of a protease is important for development of specific detection assays and selective inhibitors. In this paper we present a simplified and optimized proteomic method for the determination of protease cleavage site specificity on both prime-and non-prime side simultaneously using peptide libraries generated from whole proteomes. A a detailed step-by-step protocol for this method, including library generation and web-based data analysis, is provided.
