GraBCas: a bioinformatics tool for score-based prediction of Caspase- and Granzyme B-cleavage sites in protein sequences

Caspases and granzyme B are proteases that share the primary specificity to cleave at the carboxyl terminal of aspartate residues in their substrates. Both, caspases and granzyme B are enzymes that are involved in fundamental cellular processes and play a central role in apoptotic cell death. Although various targets are described, many substrates still await identification and many cleavage sites of known substrates are not identified or experimentally verified. A more comprehensive knowledge of caspase and granzyme B substrates is essential to understand the biological roles of these enzymes in more detail. The relatively high variability in cleavage site recognition sequence often complicates the identification of cleavage sites. As of yet there is no software available that allows identification of caspase and/or granzyme with cleavage sites differing from the consensus sequence. Here, we present a bioinformatics tool ‘GraBCas’ that provides score-based prediction of potential cleavage sites for the caspases 1–9 and granzyme B including an estimation of the fragment size. We tested GraBCas on already known substrates and showed its usefulness for protein sequence analysis. GraBCas is available at .

[1]  Alicia Algeciras-Schimnich,et al.  Apoptosis-independent functions of killer caspases. , 2002, Current opinion in cell biology.

[2]  Christoph Peters,et al.  Toward Computer-Based Cleavage Site Prediction of Cysteine Endopeptidases , 2003, Biological chemistry.

[3]  J. Lieberman,et al.  Direct cleavage of the human DNA fragmentation factor‐45 by granzyme B induces caspase‐activated DNase release and DNA fragmentation , 2001, The EMBO journal.

[4]  G. Salvesen,et al.  Internally quenched fluorescent peptide substrates disclose the subsite preferences of human caspases 1, 3, 6, 7 and 8. , 2000, The Biochemical journal.

[5]  I. H. Engels,et al.  Caspases: more than just killers? , 2001, Trends in immunology.

[6]  N. Thornberry,et al.  A Combinatorial Approach Defines Specificities of Members of the Caspase Family and Granzyme B , 1997, The Journal of Biological Chemistry.

[7]  R. Zamboni,et al.  Purification and catalytic properties of human caspase family members , 1999, Cell Death and Differentiation.

[8]  K. Schulze-Osthoff,et al.  Many cuts to ruin: a comprehensive update of caspase substrates , 2003, Cell Death and Differentiation.

[9]  J. Trapani,et al.  Initiation of Apoptosis by Granzyme B Requires Direct Cleavage of Bid, but Not Direct Granzyme B–Mediated Caspase Activation , 2000, The Journal of experimental medicine.

[10]  D. Nicholson,et al.  Activation of the apoptotic protease CPP32 by cytotoxic T-cell-derived granzyme B , 1995, Nature.

[11]  T. Ley,et al.  DFF45/ICAD can be directly processed by granzyme B during the induction of apoptosis. , 2000, Immunity.

[12]  A. Rosen,et al.  Cleavage by Granzyme B Is Strongly Predictive of Autoantigen Status , 1999, The Journal of experimental medicine.

[13]  Jennifer L. Harris,et al.  Definition and Redesign of the Extended Substrate Specificity of Granzyme B* , 1998, The Journal of Biological Chemistry.

[14]  N. Thornberry,et al.  Granzyme B directly and efficiently cleaves several downstream caspase substrates: implications for CTL-induced apoptosis. , 1998, Immunity.

[15]  Timothy J. Ley,et al.  Cytotoxic lymphocytes require granzyme B for the rapid induction of DNA fragmentation and apoptosis in allogeneic target cells , 1994, Cell.