Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases
Intestinal ischemia reperfusion injury (iIRI) is a severe clinical condition presenting high morbidity and mortality worldwide. Some of the systemic consequences of IRI can be prevented by applying ischemic preconditioning (IPC), a series of short ischemia/reperfusion events preceding the major isch...
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MDPI AG
2020-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/16/5799 |
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| author | Muhammad Tahir Samina Arshid Belchor Fontes Mariana S. Castro Simone Sidoli Veit Schwämmle Isabelle S. Luz Peter Roepstorff Wagner Fontes |
| author_facet | Muhammad Tahir Samina Arshid Belchor Fontes Mariana S. Castro Simone Sidoli Veit Schwämmle Isabelle S. Luz Peter Roepstorff Wagner Fontes |
| author_sort | Muhammad Tahir |
| collection | DOAJ |
| description | Intestinal ischemia reperfusion injury (iIRI) is a severe clinical condition presenting high morbidity and mortality worldwide. Some of the systemic consequences of IRI can be prevented by applying ischemic preconditioning (IPC), a series of short ischemia/reperfusion events preceding the major ischemia. Although neutrophils are key players in the pathophysiology of ischemic injuries, neither the dysregulation presented by these cells in iIRI nor the protective effect of iIPC have their regulation mechanisms fully understood. Protein phosphorylation, as well as the regulation of the respective phosphatases and kinases are responsible for regulating a large number of cellular functions in the inflammatory response. Moreover, in previous work we found hydrolases and transferases to be modulated in iIR and iIPC, suggesting the possible involvement of phosphatases and kinases in the process. Therefore, in the present study, we analyzed the phosphoproteome of neutrophils from rats submitted to mesenteric ischemia and reperfusion, either submitted or not to IPC, compared to quiescent controls and sham laparotomy. Proteomic analysis was performed by multi-step enrichment of phosphopeptides, isobaric labeling, and LC-MS/MS analysis. Bioinformatics was used to determine phosphosite and phosphopeptide abundance and clustering, as well as kinases and phosphatases sites and domains. We found that most of the phosphorylation-regulated proteins are involved in apoptosis and migration, and most of the regulatory kinases belong to CAMK and CMGC families. An interesting finding revealed groups of proteins that are modulated by iIR, but such modulation can be prevented by iIPC. Among the regulated proteins related to the iIPC protective effect, Vamp8 and Inpp5d/Ship are discussed as possible candidates for control of the iIR damage. |
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| last_indexed | 2024-03-10T17:32:12Z |
| publishDate | 2020-08-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-038fe6dd7a55497ca9be46557e717d322023-11-20T09:58:42ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-08-012116579910.3390/ijms21165799Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and PhosphatasesMuhammad Tahir0Samina Arshid1Belchor Fontes2Mariana S. Castro3Simone Sidoli4Veit Schwämmle5Isabelle S. Luz6Peter Roepstorff7Wagner Fontes8Laboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, Brasilia 70910-900, BrazilLaboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, Brasilia 70910-900, BrazilLaboratory of Surgical Physiopathology (LIM-62), Faculty of Medicine, University of São Paulo, São Paulo 01246903, BrazilLaboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, Brasilia 70910-900, BrazilDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, DenmarkDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, DenmarkLaboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, Brasilia 70910-900, BrazilDepartment of Biochemistry and Molecular Biology, University of Southern Denmark, DK-5230 Odense M, DenmarkLaboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, Brasilia 70910-900, BrazilIntestinal ischemia reperfusion injury (iIRI) is a severe clinical condition presenting high morbidity and mortality worldwide. Some of the systemic consequences of IRI can be prevented by applying ischemic preconditioning (IPC), a series of short ischemia/reperfusion events preceding the major ischemia. Although neutrophils are key players in the pathophysiology of ischemic injuries, neither the dysregulation presented by these cells in iIRI nor the protective effect of iIPC have their regulation mechanisms fully understood. Protein phosphorylation, as well as the regulation of the respective phosphatases and kinases are responsible for regulating a large number of cellular functions in the inflammatory response. Moreover, in previous work we found hydrolases and transferases to be modulated in iIR and iIPC, suggesting the possible involvement of phosphatases and kinases in the process. Therefore, in the present study, we analyzed the phosphoproteome of neutrophils from rats submitted to mesenteric ischemia and reperfusion, either submitted or not to IPC, compared to quiescent controls and sham laparotomy. Proteomic analysis was performed by multi-step enrichment of phosphopeptides, isobaric labeling, and LC-MS/MS analysis. Bioinformatics was used to determine phosphosite and phosphopeptide abundance and clustering, as well as kinases and phosphatases sites and domains. We found that most of the phosphorylation-regulated proteins are involved in apoptosis and migration, and most of the regulatory kinases belong to CAMK and CMGC families. An interesting finding revealed groups of proteins that are modulated by iIR, but such modulation can be prevented by iIPC. Among the regulated proteins related to the iIPC protective effect, Vamp8 and Inpp5d/Ship are discussed as possible candidates for control of the iIR damage.https://www.mdpi.com/1422-0067/21/16/5799systemic inflammatory responseischemia and reperfusionpreconditioningneutrophilsphosphorylationproteome |
| spellingShingle | Muhammad Tahir Samina Arshid Belchor Fontes Mariana S. Castro Simone Sidoli Veit Schwämmle Isabelle S. Luz Peter Roepstorff Wagner Fontes Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases International Journal of Molecular Sciences systemic inflammatory response ischemia and reperfusion preconditioning neutrophils phosphorylation proteome |
| title | Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases |
| title_full | Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases |
| title_fullStr | Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases |
| title_full_unstemmed | Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases |
| title_short | Phosphoproteomic Analysis of Rat Neutrophils Shows the Effect of Intestinal Ischemia/Reperfusion and Preconditioning on Kinases and Phosphatases |
| title_sort | phosphoproteomic analysis of rat neutrophils shows the effect of intestinal ischemia reperfusion and preconditioning on kinases and phosphatases |
| topic | systemic inflammatory response ischemia and reperfusion preconditioning neutrophils phosphorylation proteome |
| url | https://www.mdpi.com/1422-0067/21/16/5799 |
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