Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury
Background α Carboxyl terminus 1 (αCT1) is a 25–amino acid therapeutic peptide incorporating the zonula occludens‐1 (ZO‐1)–binding domain of connexin 43 (Cx43) that is currently in phase 3 clinical testing on chronic wounds. In mice, we reported that αCT1 reduced arrhythmias after cardiac injury, ac...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-08-01
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| Series: | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| Subjects: | |
| Online Access: | https://www.ahajournals.org/doi/10.1161/JAHA.119.012385 |
| _version_ | 1818315117333315584 |
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| author | Jingbo Jiang Daniel Hoagland Joseph A. Palatinus Huamei He Jegan Iyyathurai L. Jane Jourdan Geert Bultynck Zhen Wang Zhiwei Zhang Kevin Schey Steven Poelzing Francis X. McGowan Robert G. Gourdie |
| author_facet | Jingbo Jiang Daniel Hoagland Joseph A. Palatinus Huamei He Jegan Iyyathurai L. Jane Jourdan Geert Bultynck Zhen Wang Zhiwei Zhang Kevin Schey Steven Poelzing Francis X. McGowan Robert G. Gourdie |
| author_sort | Jingbo Jiang |
| collection | DOAJ |
| description | Background α Carboxyl terminus 1 (αCT1) is a 25–amino acid therapeutic peptide incorporating the zonula occludens‐1 (ZO‐1)–binding domain of connexin 43 (Cx43) that is currently in phase 3 clinical testing on chronic wounds. In mice, we reported that αCT1 reduced arrhythmias after cardiac injury, accompanied by increases in protein kinase Cε phosphorylation of Cx43 at serine 368. Herein, we characterize detailed molecular mode of action of αCT1 in mitigating cardiac ischemia‐reperfusion injury. Methods and Results To study αCT1‐mediated increases in phosphorylation of Cx43 at serine 368, we undertook mass spectrometry of protein kinase Cε phosphorylation assay reactants. This indicated potential interaction between negatively charged residues in the αCT1 Asp‐Asp‐Leu‐Glu‐Iso sequence and lysines (Lys345, Lys346) in an α‐helical sequence (helix 2) within the Cx43‐CT. In silico modeling provided further support for this interaction, indicating that αCT1 may interact with both Cx43 and ZO‐1. Using surface plasmon resonance, thermal shift, and phosphorylation assays, we characterized a series of αCT1 variants, identifying peptides that interacted with either ZO‐1–postsynaptic density‐95/disks large/zonula occludens‐1 2 or Cx43‐CT, but with limited or no ability to bind both molecules. Only peptides competent to interact with Cx43‐CT, but not ZO‐1–postsynaptic density‐95/disks large/zonula occludens‐1 2 alone, prompted increased pS368 phosphorylation. Moreover, in an ex vivo mouse model of ischemia‐reperfusion injury, preischemic infusion only with those peptides competent to bind Cx43 preserved ventricular function after ischemia‐reperfusion. Interestingly, a short 9–amino acid variant of αCT1 (αCT11) demonstrated potent cardioprotective effects when infused either before or after ischemic injury. Conclusions Interaction of αCT1 with the Cx43, but not ZO‐1, is correlated with cardioprotection. Pharmacophores targeting Cx43‐CT could provide a translational approach to preserving heart function after ischemic injury. |
| first_indexed | 2024-12-13T09:00:26Z |
| format | Article |
| id | doaj.art-6ff294b56d8a4064b96983e8ac1a14f4 |
| institution | Directory Open Access Journal |
| issn | 2047-9980 |
| language | English |
| last_indexed | 2024-12-13T09:00:26Z |
| publishDate | 2019-08-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| spelling | doaj.art-6ff294b56d8a4064b96983e8ac1a14f42022-12-21T23:53:10ZengWileyJournal of the American Heart Association: Cardiovascular and Cerebrovascular Disease2047-99802019-08-0181610.1161/JAHA.119.012385Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion InjuryJingbo Jiang0Daniel Hoagland1Joseph A. Palatinus2Huamei He3Jegan Iyyathurai4L. Jane Jourdan5Geert Bultynck6Zhen Wang7Zhiwei Zhang8Kevin Schey9Steven Poelzing10Francis X. McGowan11Robert G. Gourdie12Fralin Biomedical Research Institute at Virginia Tech Carilion Center for Heart and Reparative Medicine Research Virginia Tech Blacksburg VAFralin Biomedical Research Institute at Virginia Tech Carilion Center for Heart and Reparative Medicine Research Virginia Tech Blacksburg VACedars‐Sinai Heart Smidt Institute Cedars‐Sinai Medical Center Los Angeles CADepartment of Anesthesiology and Critical Care Medicine Children's Hospital of Philadelphia and University of Pennsylvania Philadelphia PADepartment Cellular and Molecular Medicine KU Leuven Laboratory of Molecular and Cellular Signaling Leuven BelgiumFralin Biomedical Research Institute at Virginia Tech Carilion Center for Heart and Reparative Medicine Research Virginia Tech Blacksburg VADepartment Cellular and Molecular Medicine KU Leuven Laboratory of Molecular and Cellular Signaling Leuven BelgiumDepartment of Biochemistry Vanderbilt University School of Medicine Nashville TNDepartment of Pediatric Cardiology Guangdong Cardiovascular Institute Guangdong General Hospital Guangdong Academy of Medical Sciences Guangzhou ChinaDepartment of Biochemistry Vanderbilt University School of Medicine Nashville TNFralin Biomedical Research Institute at Virginia Tech Carilion Center for Heart and Reparative Medicine Research Virginia Tech Blacksburg VADepartment of Anesthesiology and Critical Care Medicine Children's Hospital of Philadelphia and University of Pennsylvania Philadelphia PAFralin Biomedical Research Institute at Virginia Tech Carilion Center for Heart and Reparative Medicine Research Virginia Tech Blacksburg VABackground α Carboxyl terminus 1 (αCT1) is a 25–amino acid therapeutic peptide incorporating the zonula occludens‐1 (ZO‐1)–binding domain of connexin 43 (Cx43) that is currently in phase 3 clinical testing on chronic wounds. In mice, we reported that αCT1 reduced arrhythmias after cardiac injury, accompanied by increases in protein kinase Cε phosphorylation of Cx43 at serine 368. Herein, we characterize detailed molecular mode of action of αCT1 in mitigating cardiac ischemia‐reperfusion injury. Methods and Results To study αCT1‐mediated increases in phosphorylation of Cx43 at serine 368, we undertook mass spectrometry of protein kinase Cε phosphorylation assay reactants. This indicated potential interaction between negatively charged residues in the αCT1 Asp‐Asp‐Leu‐Glu‐Iso sequence and lysines (Lys345, Lys346) in an α‐helical sequence (helix 2) within the Cx43‐CT. In silico modeling provided further support for this interaction, indicating that αCT1 may interact with both Cx43 and ZO‐1. Using surface plasmon resonance, thermal shift, and phosphorylation assays, we characterized a series of αCT1 variants, identifying peptides that interacted with either ZO‐1–postsynaptic density‐95/disks large/zonula occludens‐1 2 or Cx43‐CT, but with limited or no ability to bind both molecules. Only peptides competent to interact with Cx43‐CT, but not ZO‐1–postsynaptic density‐95/disks large/zonula occludens‐1 2 alone, prompted increased pS368 phosphorylation. Moreover, in an ex vivo mouse model of ischemia‐reperfusion injury, preischemic infusion only with those peptides competent to bind Cx43 preserved ventricular function after ischemia‐reperfusion. Interestingly, a short 9–amino acid variant of αCT1 (αCT11) demonstrated potent cardioprotective effects when infused either before or after ischemic injury. Conclusions Interaction of αCT1 with the Cx43, but not ZO‐1, is correlated with cardioprotection. Pharmacophores targeting Cx43‐CT could provide a translational approach to preserving heart function after ischemic injury.https://www.ahajournals.org/doi/10.1161/JAHA.119.012385cardioprotectionconnexin 43ischemia‐reperfusion injuryS368 phosphorylationzonula occludens‐1α carboxyl terminus 1 |
| spellingShingle | Jingbo Jiang Daniel Hoagland Joseph A. Palatinus Huamei He Jegan Iyyathurai L. Jane Jourdan Geert Bultynck Zhen Wang Zhiwei Zhang Kevin Schey Steven Poelzing Francis X. McGowan Robert G. Gourdie Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease cardioprotection connexin 43 ischemia‐reperfusion injury S368 phosphorylation zonula occludens‐1 α carboxyl terminus 1 |
| title | Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury |
| title_full | Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury |
| title_fullStr | Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury |
| title_full_unstemmed | Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury |
| title_short | Interaction of α Carboxyl Terminus 1 Peptide With the Connexin 43 Carboxyl Terminus Preserves Left Ventricular Function After Ischemia‐Reperfusion Injury |
| title_sort | interaction of α carboxyl terminus 1 peptide with the connexin 43 carboxyl terminus preserves left ventricular function after ischemia reperfusion injury |
| topic | cardioprotection connexin 43 ischemia‐reperfusion injury S368 phosphorylation zonula occludens‐1 α carboxyl terminus 1 |
| url | https://www.ahajournals.org/doi/10.1161/JAHA.119.012385 |
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