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Profiling treatment‐specific post‐translational modifications in a complex proteome with subtractive substrate phage display
Author(s) -
Tenzer Angela,
Hofstetter Barbara,
Sauser Christelle,
Bodis Stephan,
Schubiger August P.,
Bonny Christophe,
Pruschy Martin
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200400811
Subject(s) - proteome , computational biology , profiling (computer programming) , phage display , subtractive color , proteomics , substrate specificity , biology , chemistry , bioinformatics , computer science , biochemistry , gene , peptide , art , visual arts , enzyme , operating system
Proteolytic activation of zymogens or controlled degradation of inhibitory factors is part of a major regulatory system on the post‐translational level to regulate treatment induced cellular stress responses. The identification of differential activity based substrates is thus of high interest to prioritize and validate candidate targets for drug discovery. Here we present a novel subtractive substrate phage display screening method for the selection of treatment induced post‐translational peptide modifications in complex proteomes. We investigated this approach with tumor cells in response to a protease activating anticancer treatment modality using subtractive and iterative screening of cellular extracts derived from control and treated cells. Specific phage were identified that served as substrates for proteolytic activities in response to treatment related activity changes and could be distinguished from substrates for unspecific proteolytic background activities. Novel, selected peptide substrates were investigated in vitro and in vivo and showed high substrate specificity and functional biological significance.