Mitochondrial p38β and Manganese Superoxide Dismutase Interaction Mediated by Estrogen in Cardiomyocytes
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{"title"=>"Mitochondrial p38β and manganese superoxide dismutase interaction mediated by estrogen in cardiomyocytes", "type"=>"journal", "authors"=>[{"first_name"=>"Han", "last_name"=>"Liu", "scopus_author_id"=>"22971738500"}, {"first_name"=>"Mounica", "last_name"=>"Yanamandala", "scopus_author_id"=>"53872301000"}, {"first_name"=>"Tiffany C.", "last_name"=>"Lee", "scopus_author_id"=>"56150884800"}, {"first_name"=>"Jin Kyung", "last_name"=>"Kim", "scopus_author_id"=>"57191681590"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"24465521", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0085272", "pui"=>"373020740", "scopus"=>"2-s2.0-84899815860", "sgr"=>"84899815860"}, "id"=>"83fcddf4-7626-35a0-a3ae-49f13ee0cbf0", "abstract"=>"AIMS: While etiology behind the observed acceleration of ischemic heart disease in postmenopausal women is poorly understood, collective scientific data suggest cardioprotective effects of the endogenous female sex hormone, estrogen. We have previously shown that 17β-estradiol (E2) protects cardiomyocytes exposed to hypoxia-reoxygenation (H/R) by inhibiting p38α - p53 signaling in apoptosis and activating pro-survival p38β mitogen activated protein kinase (p38β MAPK), leading to suppression of reactive oxygen species (ROS) post H/R. However, little is known about the mechanism behind the antioxidant actions of E2-dependent p38β. The aim of this study is to determine whether the cytoprotection by estrogen involves regulation of manganese superoxide dismutase (MnSOD), a major mitochondrial ROS scavenging enzyme, via cardiac p38β.\\n\\nMETHODS AND RESULTS: We identified mitochondrial p38β by immunocytochemistry and by immunoblotting in mitochondria isolated from neonatal cardiomyocytes of Sprague-Dawley rats. E2 facilitated the mitochondrial localization of the active form of the kinase, phosphorylated p38β (p-p38β). E2 also reduced the H/R-induced mitochondrial membrane potential decline, augmented the MnSOD activity and suppressed anion superoxide generation, while the dismutase protein expression remained unaltered. Co-immunoprecipitation studies showed physical association between MnSOD and p38β. p38β phosphorylated MnSOD in an E2-dependent manner in in-vitro kinase assays.\\n\\nCONCLUSION: This work demonstrates for the first time a mitochondrial pool of active p38β and E2-mediated phosphorylation of MnSOD by the kinase. The results shed light on the mechanism behind the cytoprotective actions of E2 in cardiomyocytes under oxidative stress.", "link"=>"http://www.mendeley.com/research/mitochondrial-p38%CE%B2-manganese-superoxide-dismutase-interaction-mediated-estrogen-cardiomyocytes", "reader_count"=>4, "reader_count_by_academic_status"=>{"Researcher"=>2, "Student > Ph. D. Student"=>1, "Other"=>1}, "reader_count_by_user_role"=>{"Researcher"=>2, "Student > Ph. D. Student"=>1, "Other"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1355424"], "description"=>"<p><i>(</i><b><i>A</i></b><i>) </i><b><i>Upper</i></b><b>,</b> Immnoblots of total p38β and phosphorylated p38β (p-p38β) from mitochondria (Mito) and cytosol (Cyto) of rat cardiomyocytes are shown. Final [E2] = 10 nM. Immunoblots of CoxIV and actin are shown also as loading controls for mitochondrial and cytosolic fractions, respectively. <b><i>Lower Left</i></b><b>,</b> CoxIV activity. Mitochondria were isolated and assayed for Cox IV activity and citrate synthase activity. CoxIV activity was then normalized to that of citrate synthase, and is presented with quantitative analysis. <b><i>Lower Right</i></b><b>,</b> NAO fluorescence. Cells were stained with NAO, a marker of mitochondrial biogenesis, and analyzed by flow cytometry. A representative graph from triplicate experiments is presented <i>(</i><b><i>B</i></b><i>)</i> Mitochondrial p-p38β. <b><i>Upper</i></b><b>,</b> the ratio of dual phosphorylated p38β (p-p38β) over total p38β in mitochondria after indicated treatments is expressed in graph. <b><i>Lower</i></b><b>,</b> the percentage of mitochondrial p38β localized to mitochondria is shown in graph as the total mitochondrial p38β over total cellular p38β pool. <i>(</i><b><i>C</i></b><i>)</i> Immunocytochemistry of p38β in cardiomyocytes. Primary antibody specific to p38β is used to stain for intracellular p38β (<i>left</i>), and the cells are co-stained with MitoTracker, a fluorescent probe for mitochondria (<i>middle</i>). The two images are then merged to demonstrate co-localization of p38β with mitochondria (<i>right</i>). The white scale bar represents 25 µm. <i>(</i><b><i>D</i></b><i>)</i> Mitochondrial membrane potential by JC-1 fluorescence. <b><i>Upper</i></b><b>,</b> Flow cytometry analysis of JC-1 stained cells after the indicated treatments shows mitochondrial membrane potential changes reflected by the ratio of the red (FL2 district) and the green (FL1) fluorescent signals. <b><i>Lower</i></b><b>,</b> Red/green JC-1 fluorescence ratio is represented in graph with quantitative analysis. * <i>p</i><0.05 vs. N, ** <i>p</i><0.05 vs. H/R.</p>", "links"=>[], "tags"=>["Biochemistry", "proteins", "Protein interactions", "hormones", "Molecular cell biology", "Signal transduction", "Signaling cascades", "Protein kinase signaling cascade", "Stress signaling cascade", "Signaling in cellular processes", "Antiapoptotic signaling", "Cellular stress responses", "cardiovascular", "Cardiovascular diseases in women", "women's health", "mitochondrial"], "article_id"=>907890, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Han Liu", "Mounica Yanamandala", "Tiffany C. Lee", "Jin Kyung Kim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0085272.g001", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_of_mitochondrial_p38_946_/907890", "title"=>"Identification of mitochondrial p38β.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-22 03:06:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1355425"], "description"=>"<p><i>(</i><b><i>A</i></b><i>)</i> p38β is immunoprecipitated from mitochondria isolated from cultured rat cardiomyocytes and subjected to kinase assays using <sup>32</sup>P-labeled ATP and ATF2 as its substrate after indicated treatments. Thus radiolabeled ATF2 is shown with quantitative analysis of the band intensity. Representative western blots of ATF2 protein and immunoprecipitated mitochondrial p38β are shown at the bottom as a control. [E2] = 10 nM. * <i>p</i><0.05 <i>vs.</i> normoxia or H/R+E2, ** <i>p</i><0.05 <i>vs.</i> H/R. <i>(</i><b><i>B</i></b><i>)</i> Viability assay. Cell viability by trypan dye exclusion is shown with the quantitative analysis with H/R (<i>left</i>) or hypoxia without reoxygenation (H) (<i>right</i>) with quantitative analysis. * <i>p</i><0.05 vs. N, ** <i>p</i><0.05 vs. H/R.</p>", "links"=>[], "tags"=>["Biochemistry", "proteins", "Protein interactions", "hormones", "Molecular cell biology", "Signal transduction", "Signaling cascades", "Protein kinase signaling cascade", "Stress signaling cascade", "Signaling in cellular processes", "Antiapoptotic signaling", "Cellular stress responses", "cardiovascular", "Cardiovascular diseases in women", "women's health"], "article_id"=>907891, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Han Liu", "Mounica Yanamandala", "Tiffany C. Lee", "Jin Kyung Kim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0085272.g002", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mitochondrial_p38_946_activity_and_viability_/907891", "title"=>"Mitochondrial p38β activity and viability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-22 03:06:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1355426"], "description"=>"<p><i>(</i><b><i>A</i></b><i>)</i> After indicated treatments, mitochondria isolated from cardiomyocytes are assayed for MnSOD activity. The final dismutase activity is expressed relative to that under normoxia (control), with quantitative analysis. * <i>p</i><0.05 <i>vs.</i> H/R. ** <i>p</i><0.05 <i>vs.</i> H/R+E2, H/R+PPT and H/R+DPN. Western blots of the MnSOD protein and CoxIV are shown below for controls. The final concentration of E2, PPT, and DPN was10 nM, and 100 nM for the ER inhibitors, MPP and PHTPP. <i>(</i><b><i>B</i></b><i>)</i> Anion superoxide assay. Intracellular anion superoxide production is determined by MitoSOX™ Red mitochondrial superoxide indicator fluorescence after the indicated treatments and various E2 final concentrations, and presented in graph with quantitative analysis. * <i>p</i><0.05 <i>vs.</i> N, ** <i>p</i><0.05 <i>vs.</i> H/R, # <i>p</i><0.05 <i>vs.</i> H/R, H/R+E2[0.1 nM] or H/R+E2[1 µM]. <i>(</i><b><i>C</i></b><i>)</i> MnSOD activity (<i>left</i>) and mito p38β translocation (<i>right</i>) after hypoxia without reoxygenation (H) are presented in graph with quantitative analysis. * <i>p</i><0.05 <i>vs.</i> N. Western blots of MnSOD, CoxIV protein, and actin are shown for loading control.</p>", "links"=>[], "tags"=>["Biochemistry", "proteins", "Protein interactions", "hormones", "Molecular cell biology", "Signal transduction", "Signaling cascades", "Protein kinase signaling cascade", "Stress signaling cascade", "Signaling in cellular processes", "Antiapoptotic signaling", "Cellular stress responses", "cardiovascular", "Cardiovascular diseases in women", "women's health", "anion"], "article_id"=>907892, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Han Liu", "Mounica Yanamandala", "Tiffany C. Lee", "Jin Kyung Kim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0085272.g003", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MnSOD_activity_and_anion_superoxide_/907892", "title"=>"MnSOD activity and anion superoxide.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-22 03:06:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1355427"], "description"=>"<p><i>(</i><b><i>A</i></b><i>)</i> Endogenous p38β is immunoprecipitated (IP: p38β) from cardiomyocytes after H/R with or without E2 at 10 nM, and the p38β-containing immunocomplex blotted for MnSOD. A western blot of p38β protein is also shown as a loading control. Nonspecific IgG is used as a specificity control in place of antibody specific to p38β in immunoprecipitation (IP: NS IgG). <i>(</i><b><i>B</i></b><i>)</i> Endogenous MnSOD is immunoprecipitated (IP: MnSOD) from treated cardiomyocytes as indicated, then the MnSOD-containing immunocomplex is blotted for p38β. Nonspecific IgG is used as a specificity control in place of antibody specific to MnSOD in immunoprecipitation (IP: NS IgG).</p>", "links"=>[], "tags"=>["Biochemistry", "proteins", "Protein interactions", "hormones", "Molecular cell biology", "Signal transduction", "Signaling cascades", "Protein kinase signaling cascade", "Stress signaling cascade", "Signaling in cellular processes", "Antiapoptotic signaling", "Cellular stress responses", "cardiovascular", "Cardiovascular diseases in women", "women's health"], "article_id"=>907893, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Han Liu", "Mounica Yanamandala", "Tiffany C. Lee", "Jin Kyung Kim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0085272.g004", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Physical_association_between_p38_946_and_MnSOD_/907893", "title"=>"Physical association between p38β and MnSOD.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-22 03:06:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/1355428"], "description"=>"<p><i>(</i><b><i>A</i></b><i>)</i> In-vitro kinase assay is performed with purified MnSOD in the indicated amount as a substrate in the presence of activated, GST-tagged purified p38β kinase in the presence or absence of the specific kinase inhibitor, SB203580. As a positive control, the kinase activity is also assayed on a well known substrate, ATF2. <i>(</i><b><i>B</i></b><i>)</i> In-vitro kinase assay is performed with endogenous p38β kinase immunoprecipitated from cultured rat cardiomyocytes and 1 µg of MnSOD or ATF2 as substrate in the presence or absence of the kinase inhibitor. A western blot of the isolated kinase is shown as a control.</p>", "links"=>[], "tags"=>["Biochemistry", "proteins", "Protein interactions", "hormones", "Molecular cell biology", "Signal transduction", "Signaling cascades", "Protein kinase signaling cascade", "Stress signaling cascade", "Signaling in cellular processes", "Antiapoptotic signaling", "Cellular stress responses", "cardiovascular", "Cardiovascular diseases in women", "women's health", "mnsod"], "article_id"=>907894, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Han Liu", "Mounica Yanamandala", "Tiffany C. Lee", "Jin Kyung Kim"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0085272.g005", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phosphorylation_of_MnSOD_by_p38_946_kinase_/907894", "title"=>"Phosphorylation of MnSOD by p38β kinase.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-01-22 03:06:57"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[291]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[286]}, {"subject_area"=>"/Biology and life sciences/Physiology", "average_usage"=>[280]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[285]}, {"subject_area"=>"/Medicine and health sciences/Physiology", "average_usage"=>[278]}]}
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