Title: Proteome-wide substrate analysis indicates substrate exclusion as a mechanism to generate caspase-7 versus caspase-3 specificity
Authors: Demon, Dieter ×
Van Damme, Petra
Vanden Berghe, Tom
Deceuninck, Annelies
Van Durme, Joost
Verspurten, Jelle
Helsens, Kenny
Impens, Francis
Wejda, Magdalena
Schymkowitz, Joost
Rousseau, Frederic
Madder, Annemieke
Vandekerckhove, Joël
Declercq, Wim
Gevaert, Kris
Vandenabeele, Peter #
Issue Date: Dec-2009
Publisher: American Society for Biochemistry and Molecular Biology
Series Title: Molecular & Cellular Proteomics vol:8 issue:12 pages:2700-2714
Abstract: Caspase-3 and -7 are considered functionally redundant proteases with similar proteolytic specificities. We performed a proteome-wide screen on a mouse macrophage lysate using the N-terminal combined fractional diagonal chromatography technology and identified 46 shared, three caspase-3-specific, and six caspase-7-specific cleavage sites. Further analysis of these cleavage sites and substitution mutation experiments revealed that for certain cleavage sites a lysine at the P5 position contributes to the discrimination between caspase-7 and -3 specificity. One of the caspase-7-specific substrates, the 40 S ribosomal protein S18, was studied in detail. The RPS18-derived P6-P5' undecapeptide retained complete specificity for caspase-7. The corresponding P6-P1 hexapeptide still displayed caspase-7 preference but lost strict specificity, suggesting that P' residues are additionally required for caspase-7-specific cleavage. Analysis of truncated peptide mutants revealed that in the case of RPS18 the P4-P1 residues constitute the core cleavage site but that P6, P5, P2', and P3' residues critically contribute to caspase-7 specificity. Interestingly, specific cleavage by caspase-7 relies on excluding recognition by caspase-3 and not on increasing binding for caspase-7.
ISSN: 1535-9476
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Non-KU Leuven Association publications
× corresponding author
# (joint) last author

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