Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
Background Arginase II activity contributes to reciprocal regulation of endothelial nitric oxide synthase (eNOS). We tested the hypotheses that arginase II activity participates in the regulation of Ca2+/Ca2+/calmodulin‐dependent kinase II/eNOS activation, and this process is dependent on mitochondr...
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| Format: | Article |
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Wiley
2018-09-01
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| Series: | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
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| Online Access: | https://www.ahajournals.org/doi/10.1161/JAHA.118.009579 |
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| author | Bon‐Hyeock Koo Hye‐Mi Hwang Bong‐Gu Yi Hyun Kyo Lim Byeong Hwa Jeon Kwang Lae Hoe Young‐Guen Kwon Moo‐Ho Won Young Myeong Kim Dan E. Berkowitz Sungwoo Ryoo |
| author_facet | Bon‐Hyeock Koo Hye‐Mi Hwang Bong‐Gu Yi Hyun Kyo Lim Byeong Hwa Jeon Kwang Lae Hoe Young‐Guen Kwon Moo‐Ho Won Young Myeong Kim Dan E. Berkowitz Sungwoo Ryoo |
| author_sort | Bon‐Hyeock Koo |
| collection | DOAJ |
| description | Background Arginase II activity contributes to reciprocal regulation of endothelial nitric oxide synthase (eNOS). We tested the hypotheses that arginase II activity participates in the regulation of Ca2+/Ca2+/calmodulin‐dependent kinase II/eNOS activation, and this process is dependent on mitochondrial p32. Methods and Results Downregulation of arginase II increased the concentration of cytosolic Ca2+ ([Ca2+]c) and decreased mitochondrial Ca2+ ([Ca2+]m) in microscopic and fluorescence‐activated cell sorting analyses, resulting in augmented eNOS Ser1177 phosphorylation and decreased eNOS Thr495 phosphorylation through Ca2+/Ca2+/calmodulin‐dependent kinase II. These changes were observed in human umbilical vein endothelial cells treated with small interfering RNA against p32 (sip32). Using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry, fluorescence immunoassay, and ion chromatography, inhibition of arginase II reduced the amount of spermine, a binding molecule, and the release of Ca2+ from p32. In addition, arginase II gene knockdown using small interfering RNA and knockout arginase II‐null mice resulted in reduced p32 protein level. In the aortas of wild‐type mice, small interfering RNA against p32 induced eNOS Ser1177 phosphorylation and enhanced NO‐dependent vasorelaxation. Arginase activity, p32 protein expression, spermine amount, and [Ca2+]m were increased in the aortas from apolipoprotein E (ApoE−/−) mice fed a high‐cholesterol diet, and intravenous administration of small interfering RNA against p32 restored Ca2+/Ca2+/calmodulin‐dependent kinase II‐dependent eNOS Ser1177 phosphorylation and improved endothelial dysfunction. The effects of arginase II downregulation were not associated with elevated NO production when tested in aortic endothelia from eNOS knockout mice. Conclusions These data demonstrate a novel function of arginase II in regulation of Ca2+‐dependent eNOS phosphorylation. This novel mechanism drives arginase activation, mitochondrial dysfunction, endothelial dysfunction, and atherogenesis. |
| format | Article |
| id | doaj-art-1dae1012683042eebada206f0a47a565 |
| institution | DOAJ |
| issn | 2047-9980 |
| language | English |
| publishDate | 2018-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| spelling | doaj-art-1dae1012683042eebada206f0a47a5652025-08-20T03:07:46ZengWileyJournal of the American Heart Association: Cardiovascular and Cerebrovascular Disease2047-99802018-09-0171810.1161/JAHA.118.009579Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent MannerBon‐Hyeock Koo0Hye‐Mi Hwang1Bong‐Gu Yi2Hyun Kyo Lim3Byeong Hwa Jeon4Kwang Lae Hoe5Young‐Guen Kwon6Moo‐Ho Won7Young Myeong Kim8Dan E. Berkowitz9Sungwoo Ryoo10Department of Biology School of medicine Kangwon National University Chuncheon KoreaDepartment of Biology School of medicine Kangwon National University Chuncheon KoreaDepartment of Biology School of medicine Kangwon National University Chuncheon KoreaDepartment of Anesthesiology and Pain Medicine Yonsei University Wonju College of Medicine Wonju KoreaInfectious Signaling Network Research Center Department of Physiology School of Medicine Chungnam National University Daejeon KoreaDepartment of New Drug Discovery and Development Chungnam National University Daejeon KoreaDepartment of Biochemistry Yonsei University Seoul KoreaDepartment of Neurobiology School of medicine Kangwon National University Chuncheon KoreaCollege of Natural Sciences and Departments of Molecular and Cellular Biochemistry School of medicine Kangwon National University Chuncheon KoreaDepartment of Anesthesiology and Critical Care Medicine Johns Hopkins University Baltimore MDDepartment of Biology School of medicine Kangwon National University Chuncheon KoreaBackground Arginase II activity contributes to reciprocal regulation of endothelial nitric oxide synthase (eNOS). We tested the hypotheses that arginase II activity participates in the regulation of Ca2+/Ca2+/calmodulin‐dependent kinase II/eNOS activation, and this process is dependent on mitochondrial p32. Methods and Results Downregulation of arginase II increased the concentration of cytosolic Ca2+ ([Ca2+]c) and decreased mitochondrial Ca2+ ([Ca2+]m) in microscopic and fluorescence‐activated cell sorting analyses, resulting in augmented eNOS Ser1177 phosphorylation and decreased eNOS Thr495 phosphorylation through Ca2+/Ca2+/calmodulin‐dependent kinase II. These changes were observed in human umbilical vein endothelial cells treated with small interfering RNA against p32 (sip32). Using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry, fluorescence immunoassay, and ion chromatography, inhibition of arginase II reduced the amount of spermine, a binding molecule, and the release of Ca2+ from p32. In addition, arginase II gene knockdown using small interfering RNA and knockout arginase II‐null mice resulted in reduced p32 protein level. In the aortas of wild‐type mice, small interfering RNA against p32 induced eNOS Ser1177 phosphorylation and enhanced NO‐dependent vasorelaxation. Arginase activity, p32 protein expression, spermine amount, and [Ca2+]m were increased in the aortas from apolipoprotein E (ApoE−/−) mice fed a high‐cholesterol diet, and intravenous administration of small interfering RNA against p32 restored Ca2+/Ca2+/calmodulin‐dependent kinase II‐dependent eNOS Ser1177 phosphorylation and improved endothelial dysfunction. The effects of arginase II downregulation were not associated with elevated NO production when tested in aortic endothelia from eNOS knockout mice. Conclusions These data demonstrate a novel function of arginase II in regulation of Ca2+‐dependent eNOS phosphorylation. This novel mechanism drives arginase activation, mitochondrial dysfunction, endothelial dysfunction, and atherogenesis.https://www.ahajournals.org/doi/10.1161/JAHA.118.009579Arginase IIatherogenesisCalcium signalingeNOSp32 |
| spellingShingle | Bon‐Hyeock Koo Hye‐Mi Hwang Bong‐Gu Yi Hyun Kyo Lim Byeong Hwa Jeon Kwang Lae Hoe Young‐Guen Kwon Moo‐Ho Won Young Myeong Kim Dan E. Berkowitz Sungwoo Ryoo Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease Arginase II atherogenesis Calcium signaling eNOS p32 |
| title | Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner |
| title_full | Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner |
| title_fullStr | Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner |
| title_full_unstemmed | Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner |
| title_short | Arginase II Contributes to the Ca2+/CaMKII/eNOS Axis by Regulating Ca2+ Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner |
| title_sort | arginase ii contributes to the ca2 camkii enos axis by regulating ca2 concentration between the cytosol and mitochondria in a p32 dependent manner |
| topic | Arginase II atherogenesis Calcium signaling eNOS p32 |
| url | https://www.ahajournals.org/doi/10.1161/JAHA.118.009579 |
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