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Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury
As an integral component of cell membrane repair machinery, MG53 (mitsugumin 53) is important for cardioprotection induced by ischemia preconditioning and postconditioning. However, it also impairs insulin signaling via its E3 ligase activity-mediated ubiquitination-dependent degradation of IR (insu...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Lippincott Williams & Wilkins
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9770150/ https://www.ncbi.nlm.nih.gov/pubmed/36337049 http://dx.doi.org/10.1161/CIRCRESAHA.122.321055 |
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author | Lv, Fengxiang Wang, Yingfan Shan, Dan Guo, Sile Chen, Gengjia Jin, Li Zheng, Wen Feng, Han Zeng, Xiaohu Zhang, Shuo Zhang, Yan Hu, Xinli Xiao, Rui-Ping |
author_facet | Lv, Fengxiang Wang, Yingfan Shan, Dan Guo, Sile Chen, Gengjia Jin, Li Zheng, Wen Feng, Han Zeng, Xiaohu Zhang, Shuo Zhang, Yan Hu, Xinli Xiao, Rui-Ping |
author_sort | Lv, Fengxiang |
collection | PubMed |
description | As an integral component of cell membrane repair machinery, MG53 (mitsugumin 53) is important for cardioprotection induced by ischemia preconditioning and postconditioning. However, it also impairs insulin signaling via its E3 ligase activity-mediated ubiquitination-dependent degradation of IR (insulin receptor) and IRS1 (insulin receptor substrate 1) and its myokine function-induced allosteric blockage of IR. Here, we sought to develop MG53 into a cardioprotection therapy by separating its detrimental metabolic effects from beneficial actions. METHODS: Using immunoprecipitation-mass spectrometry, site-specific mutation, in vitro kinase assay, and in vivo animal studies, we investigated the role of MG53 phosphorylation at serine 255 (S255). In particular, utilizing recombinant proteins and gene knock-in approaches, we evaluated the potential therapeutic effect of MG53-S255A mutant in treating cardiac ischemia/reperfusion injury in diabetic mice. RESULTS: We identified S255 phosphorylation as a prerequisite for MG53 E3 ligase activity. Furthermore, MG53(S255) phosphorylation was mediated by GSK3β (glycogen synthase kinase 3 beta) and markedly elevated in the animal models with metabolic disorders. Thus, IR-IRS1-GSK3β-MG53 formed a vicious cycle in the pathogenesis of metabolic disorders where aberrant insulin signaling led to hyper-activation of GSK3β, which in turn, phosphorylated MG53 and enhanced its E3 ligase activity, and further impaired insulin sensitivity. Importantly, S255A mutant eliminated the E3 ligase activity while retained cell protective function of MG53. Consequently, the S255A mutant, but not the wild type MG53, protected the heart against ischemia/reperfusion injury in db/db mice with advanced diabetes, although both elicited cardioprotection in normal mice. Moreover, in S255A knock-in mice, S255A mutant also mitigated ischemia/reperfusion-induced myocardial damage in the diabetic setting. CONCLUSIONS: S255 phosphorylation is a biased regulation of MG53 E3 ligase activity. The MG53-S255A mutant provides a promising approach for the treatment of acute myocardial injury, especially in patients with metabolic disorders. |
format | Online Article Text |
id | pubmed-9770150 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Lippincott Williams & Wilkins |
record_format | MEDLINE/PubMed |
spelling | pubmed-97701502022-12-28 Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury Lv, Fengxiang Wang, Yingfan Shan, Dan Guo, Sile Chen, Gengjia Jin, Li Zheng, Wen Feng, Han Zeng, Xiaohu Zhang, Shuo Zhang, Yan Hu, Xinli Xiao, Rui-Ping Circ Res Original Research As an integral component of cell membrane repair machinery, MG53 (mitsugumin 53) is important for cardioprotection induced by ischemia preconditioning and postconditioning. However, it also impairs insulin signaling via its E3 ligase activity-mediated ubiquitination-dependent degradation of IR (insulin receptor) and IRS1 (insulin receptor substrate 1) and its myokine function-induced allosteric blockage of IR. Here, we sought to develop MG53 into a cardioprotection therapy by separating its detrimental metabolic effects from beneficial actions. METHODS: Using immunoprecipitation-mass spectrometry, site-specific mutation, in vitro kinase assay, and in vivo animal studies, we investigated the role of MG53 phosphorylation at serine 255 (S255). In particular, utilizing recombinant proteins and gene knock-in approaches, we evaluated the potential therapeutic effect of MG53-S255A mutant in treating cardiac ischemia/reperfusion injury in diabetic mice. RESULTS: We identified S255 phosphorylation as a prerequisite for MG53 E3 ligase activity. Furthermore, MG53(S255) phosphorylation was mediated by GSK3β (glycogen synthase kinase 3 beta) and markedly elevated in the animal models with metabolic disorders. Thus, IR-IRS1-GSK3β-MG53 formed a vicious cycle in the pathogenesis of metabolic disorders where aberrant insulin signaling led to hyper-activation of GSK3β, which in turn, phosphorylated MG53 and enhanced its E3 ligase activity, and further impaired insulin sensitivity. Importantly, S255A mutant eliminated the E3 ligase activity while retained cell protective function of MG53. Consequently, the S255A mutant, but not the wild type MG53, protected the heart against ischemia/reperfusion injury in db/db mice with advanced diabetes, although both elicited cardioprotection in normal mice. Moreover, in S255A knock-in mice, S255A mutant also mitigated ischemia/reperfusion-induced myocardial damage in the diabetic setting. CONCLUSIONS: S255 phosphorylation is a biased regulation of MG53 E3 ligase activity. The MG53-S255A mutant provides a promising approach for the treatment of acute myocardial injury, especially in patients with metabolic disorders. Lippincott Williams & Wilkins 2022-11-07 2022-12-02 /pmc/articles/PMC9770150/ /pubmed/36337049 http://dx.doi.org/10.1161/CIRCRESAHA.122.321055 Text en © 2022 The Authors. https://creativecommons.org/licenses/by/4.0/Circulation Research is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited. |
spellingShingle | Original Research Lv, Fengxiang Wang, Yingfan Shan, Dan Guo, Sile Chen, Gengjia Jin, Li Zheng, Wen Feng, Han Zeng, Xiaohu Zhang, Shuo Zhang, Yan Hu, Xinli Xiao, Rui-Ping Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title | Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title_full | Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title_fullStr | Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title_full_unstemmed | Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title_short | Blocking MG53(S255) Phosphorylation Protects Diabetic Heart From Ischemic Injury |
title_sort | blocking mg53(s255) phosphorylation protects diabetic heart from ischemic injury |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9770150/ https://www.ncbi.nlm.nih.gov/pubmed/36337049 http://dx.doi.org/10.1161/CIRCRESAHA.122.321055 |
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