Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes
In the heart, alternative splicing of the igf-I gene produces two isoforms: IGF-IEa and IGF-IEc, (Mechano-growth factor, MGF). The sequence divergence between their E-domain regions suggests differential isoform function. To define the biological actions of MGF’s E-domain, we performed in silico ana...
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| Format: | Article |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Physiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphys.2022.1028345/full |
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| author | Christopher Solís Walter C. Thompson James R. Peña Christopher McDermott-Roe Paulina Langa Paulina Langa Chad M. Warren Chad M. Warren Magdalena Chrzanowska Beata M. Wolska Beata M. Wolska R. John Solaro R. John Solaro Pieter Detombe Pieter Detombe Paul H. Goldspink Paul H. Goldspink |
| author_facet | Christopher Solís Walter C. Thompson James R. Peña Christopher McDermott-Roe Paulina Langa Paulina Langa Chad M. Warren Chad M. Warren Magdalena Chrzanowska Beata M. Wolska Beata M. Wolska R. John Solaro R. John Solaro Pieter Detombe Pieter Detombe Paul H. Goldspink Paul H. Goldspink |
| author_sort | Christopher Solís |
| collection | DOAJ |
| description | In the heart, alternative splicing of the igf-I gene produces two isoforms: IGF-IEa and IGF-IEc, (Mechano-growth factor, MGF). The sequence divergence between their E-domain regions suggests differential isoform function. To define the biological actions of MGF’s E-domain, we performed in silico analysis of the unique C-terminal sequence and identified a phosphorylation consensus site residing within a putative 14-3-3 binding motif. To test the functional significance of Ser 18 phosphorylation, phospho-mimetic (S/E18) and phospho-null (S/A18) peptides were delivered to mice at different doses for 2 weeks. Cardiovascular function was measured using echocardiography and a pressure-volume catheter. At the lowest (2.25 mg/kg/day) and highest (9 mg/kg/day) doses, the peptides produced a depression in systolic and diastolic parameters. However, at 4.5 mg/kg/day the peptides produced opposing effects on cardiac function. Fractional shortening analysis also showed a similar trend, but with no significant change in cardiac geometry. Microarray analysis discovered 21 genes (FDR p < 0.01), that were expressed accordant with the opposing effects on contractile function at 4.5 mg/kg/day, with the nuclear receptor subfamily 4 group A member 2 (Nr4a2) identified as a potential target of peptide regulation. Testing the regulation of the Nr4a family, showed the E-domain peptides modulate Nr4a gene expression following membrane depolarization with KCl in vitro. To determine the potential role of 14-3-3 proteins, we examined 14-3-3 isoform expression and distribution. 14-3-3γ localized to the myofilaments in neonatal cardiac myocytes, the cardiac myocytes and myofilament extracts from the adult heart. Thermal shift analysis of recombinant 14-3-3γ protein showed the S/A18 peptide destabilized 14-3-3γ folding. Also, the S/A18 peptide significantly inhibited 14-3-3γ’s ability to interact with myosin binding protein C (MYPC3) and phospholamban (PLN) in heart lysates from dobutamine injected mice. Conversely, the S/E18 peptide showed no effect on 14-3-3γ stability, did not inhibit 14-3-3γ’s interaction with PLN but did inhibit the interaction with MYPC3. Replacing the glutamic acid with a phosphate group on Ser 18 (pSer18), significantly increased 14-3-3γ protein stability. We conclude that the state of Ser 18 phosphorylation within the 14-3-3 binding motif of MGF’s E-domain, modulates protein-protein interactions within the 14-3-3γ interactome, which includes proteins involved in the regulation of contractile function. |
| first_indexed | 2024-04-13T11:00:53Z |
| format | Article |
| id | doaj.art-4276e5406b9d47d2af2e397c632f998a |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-04-13T11:00:53Z |
| publishDate | 2022-11-01 |
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| series | Frontiers in Physiology |
| spelling | doaj.art-4276e5406b9d47d2af2e397c632f998a2022-12-22T02:49:25ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2022-11-011310.3389/fphys.2022.10283451028345Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomesChristopher Solís0Walter C. Thompson1James R. Peña2Christopher McDermott-Roe3Paulina Langa4Paulina Langa5Chad M. Warren6Chad M. Warren7Magdalena Chrzanowska8Beata M. Wolska9Beata M. Wolska10R. John Solaro11R. John Solaro12 Pieter Detombe13 Pieter Detombe14Paul H. Goldspink15Paul H. Goldspink16Department of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesDepartment of Physiology, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Medicine, and Department of Genetics, Perelman School of Medicine, Cardiovascular Institute, University of Pennsylvania, Philadelphia, PA, United StatesDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesCenter for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesCenter for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, United StatesBlood Research Institute, Versiti, Department of Pharmacology and Toxicology, Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI, United StatesCenter for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Medicine, Division of Cardiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesDepartment of Physiology, Medical College of Wisconsin, Milwaukee, WI, United StatesDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesPhymedexp, Université de Montpellier, Inserm, CNRS, Montpellier, FranceDepartment of Physiology and Biophysics, University of Illinois Chicago, Chicago, IL, United StatesDepartment of Physiology, Medical College of Wisconsin, Milwaukee, WI, United StatesIn the heart, alternative splicing of the igf-I gene produces two isoforms: IGF-IEa and IGF-IEc, (Mechano-growth factor, MGF). The sequence divergence between their E-domain regions suggests differential isoform function. To define the biological actions of MGF’s E-domain, we performed in silico analysis of the unique C-terminal sequence and identified a phosphorylation consensus site residing within a putative 14-3-3 binding motif. To test the functional significance of Ser 18 phosphorylation, phospho-mimetic (S/E18) and phospho-null (S/A18) peptides were delivered to mice at different doses for 2 weeks. Cardiovascular function was measured using echocardiography and a pressure-volume catheter. At the lowest (2.25 mg/kg/day) and highest (9 mg/kg/day) doses, the peptides produced a depression in systolic and diastolic parameters. However, at 4.5 mg/kg/day the peptides produced opposing effects on cardiac function. Fractional shortening analysis also showed a similar trend, but with no significant change in cardiac geometry. Microarray analysis discovered 21 genes (FDR p < 0.01), that were expressed accordant with the opposing effects on contractile function at 4.5 mg/kg/day, with the nuclear receptor subfamily 4 group A member 2 (Nr4a2) identified as a potential target of peptide regulation. Testing the regulation of the Nr4a family, showed the E-domain peptides modulate Nr4a gene expression following membrane depolarization with KCl in vitro. To determine the potential role of 14-3-3 proteins, we examined 14-3-3 isoform expression and distribution. 14-3-3γ localized to the myofilaments in neonatal cardiac myocytes, the cardiac myocytes and myofilament extracts from the adult heart. Thermal shift analysis of recombinant 14-3-3γ protein showed the S/A18 peptide destabilized 14-3-3γ folding. Also, the S/A18 peptide significantly inhibited 14-3-3γ’s ability to interact with myosin binding protein C (MYPC3) and phospholamban (PLN) in heart lysates from dobutamine injected mice. Conversely, the S/E18 peptide showed no effect on 14-3-3γ stability, did not inhibit 14-3-3γ’s interaction with PLN but did inhibit the interaction with MYPC3. Replacing the glutamic acid with a phosphate group on Ser 18 (pSer18), significantly increased 14-3-3γ protein stability. We conclude that the state of Ser 18 phosphorylation within the 14-3-3 binding motif of MGF’s E-domain, modulates protein-protein interactions within the 14-3-3γ interactome, which includes proteins involved in the regulation of contractile function.https://www.frontiersin.org/articles/10.3389/fphys.2022.1028345/fullmechano-growth factorE-domain peptide14-3-3 proteincontractile function of the heartprotein-protein interaction (PPI) |
| spellingShingle | Christopher Solís Walter C. Thompson James R. Peña Christopher McDermott-Roe Paulina Langa Paulina Langa Chad M. Warren Chad M. Warren Magdalena Chrzanowska Beata M. Wolska Beata M. Wolska R. John Solaro R. John Solaro Pieter Detombe Pieter Detombe Paul H. Goldspink Paul H. Goldspink Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes Frontiers in Physiology mechano-growth factor E-domain peptide 14-3-3 protein contractile function of the heart protein-protein interaction (PPI) |
| title | Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes |
| title_full | Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes |
| title_fullStr | Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes |
| title_full_unstemmed | Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes |
| title_short | Mechano-growth factor E-domain modulates cardiac contractile function through 14-3-3 protein interactomes |
| title_sort | mechano growth factor e domain modulates cardiac contractile function through 14 3 3 protein interactomes |
| topic | mechano-growth factor E-domain peptide 14-3-3 protein contractile function of the heart protein-protein interaction (PPI) |
| url | https://www.frontiersin.org/articles/10.3389/fphys.2022.1028345/full |
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