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An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation
Post-translational modifications of proteins regulate diverse cellular functions, with mounting evidence suggesting that hierarchical cross-talk between distinct modifications may fine-tune cellular responses. For example, in apoptosis, caspases promote cell death via cleavage of key structural and...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Biochemistry and Molecular Biology
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014278/ https://www.ncbi.nlm.nih.gov/pubmed/24556848 http://dx.doi.org/10.1074/mcp.M113.037374 |
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author | Turowec, Jacob P. Zukowski, Stephanie A. Knight, James D. R. Smalley, David M. Graves, Lee M. Johnson, Gary L. Li, Shawn S. C. Lajoie, Gilles A. Litchfield, David W. |
author_facet | Turowec, Jacob P. Zukowski, Stephanie A. Knight, James D. R. Smalley, David M. Graves, Lee M. Johnson, Gary L. Li, Shawn S. C. Lajoie, Gilles A. Litchfield, David W. |
author_sort | Turowec, Jacob P. |
collection | PubMed |
description | Post-translational modifications of proteins regulate diverse cellular functions, with mounting evidence suggesting that hierarchical cross-talk between distinct modifications may fine-tune cellular responses. For example, in apoptosis, caspases promote cell death via cleavage of key structural and enzymatic proteins that in some instances is inhibited by phosphorylation near the scissile bond. In this study, we systematically investigated how protein phosphorylation affects susceptibility to caspase cleavage using an N-terminomic strategy, namely, a modified terminal amino isotopic labeling of substrates (TAILS) workflow, to identify proteins for which caspase-catalyzed cleavage is modulated by phosphatase treatment. We validated the effects of phosphorylation on three of the identified proteins and found that Yap1 and Golgin-160 exhibit decreased cleavage when phosphorylated, whereas cleavage of MST3 was promoted by phosphorylation. Furthermore, using synthetic peptides we systematically examined the influence of phosphoserine throughout the entirety of caspase-3, -7, and -8 recognition motifs and observed a general inhibitory effect of phosphorylation even at residues considered outside the classical consensus motif. Overall, our work demonstrates a role for phosphorylation in controlling caspase-mediated cleavage and shows that N-terminomic strategies can be tailored to study cross-talk between phosphorylation and proteolysis. |
format | Online Article Text |
id | pubmed-4014278 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | The American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-40142782014-05-09 An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation Turowec, Jacob P. Zukowski, Stephanie A. Knight, James D. R. Smalley, David M. Graves, Lee M. Johnson, Gary L. Li, Shawn S. C. Lajoie, Gilles A. Litchfield, David W. Mol Cell Proteomics Research Post-translational modifications of proteins regulate diverse cellular functions, with mounting evidence suggesting that hierarchical cross-talk between distinct modifications may fine-tune cellular responses. For example, in apoptosis, caspases promote cell death via cleavage of key structural and enzymatic proteins that in some instances is inhibited by phosphorylation near the scissile bond. In this study, we systematically investigated how protein phosphorylation affects susceptibility to caspase cleavage using an N-terminomic strategy, namely, a modified terminal amino isotopic labeling of substrates (TAILS) workflow, to identify proteins for which caspase-catalyzed cleavage is modulated by phosphatase treatment. We validated the effects of phosphorylation on three of the identified proteins and found that Yap1 and Golgin-160 exhibit decreased cleavage when phosphorylated, whereas cleavage of MST3 was promoted by phosphorylation. Furthermore, using synthetic peptides we systematically examined the influence of phosphoserine throughout the entirety of caspase-3, -7, and -8 recognition motifs and observed a general inhibitory effect of phosphorylation even at residues considered outside the classical consensus motif. Overall, our work demonstrates a role for phosphorylation in controlling caspase-mediated cleavage and shows that N-terminomic strategies can be tailored to study cross-talk between phosphorylation and proteolysis. The American Society for Biochemistry and Molecular Biology 2014-05 2014-02-20 /pmc/articles/PMC4014278/ /pubmed/24556848 http://dx.doi.org/10.1074/mcp.M113.037374 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. |
spellingShingle | Research Turowec, Jacob P. Zukowski, Stephanie A. Knight, James D. R. Smalley, David M. Graves, Lee M. Johnson, Gary L. Li, Shawn S. C. Lajoie, Gilles A. Litchfield, David W. An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title | An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title_full | An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title_fullStr | An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title_full_unstemmed | An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title_short | An Unbiased Proteomic Screen Reveals Caspase Cleavage Is Positively and Negatively Regulated by Substrate Phosphorylation |
title_sort | unbiased proteomic screen reveals caspase cleavage is positively and negatively regulated by substrate phosphorylation |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014278/ https://www.ncbi.nlm.nih.gov/pubmed/24556848 http://dx.doi.org/10.1074/mcp.M113.037374 |
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