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Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) 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 Ca(2+)/Ca(2+)/calmodulin‐dependent kinase II/eNOS activation, and this process is dependent on mitoc...

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Autores principales: Koo, Bon‐Hyeock, Hwang, Hye‐Mi, Yi, Bong‐Gu, Lim, Hyun Kyo, Jeon, Byeong Hwa, Hoe, Kwang Lae, Kwon, Young‐Guen, Won, Moo‐Ho, Kim, Young Myeong, Berkowitz, Dan E., Ryoo, Sungwoo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6222941/
https://www.ncbi.nlm.nih.gov/pubmed/30371203
http://dx.doi.org/10.1161/JAHA.118.009579
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author Koo, Bon‐Hyeock
Hwang, Hye‐Mi
Yi, Bong‐Gu
Lim, Hyun Kyo
Jeon, Byeong Hwa
Hoe, Kwang Lae
Kwon, Young‐Guen
Won, Moo‐Ho
Kim, Young Myeong
Berkowitz, Dan E.
Ryoo, Sungwoo
author_facet Koo, Bon‐Hyeock
Hwang, Hye‐Mi
Yi, Bong‐Gu
Lim, Hyun Kyo
Jeon, Byeong Hwa
Hoe, Kwang Lae
Kwon, Young‐Guen
Won, Moo‐Ho
Kim, Young Myeong
Berkowitz, Dan E.
Ryoo, Sungwoo
author_sort Koo, Bon‐Hyeock
collection PubMed
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 Ca(2+)/Ca(2+)/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 Ca(2+) ([Ca(2+)]c) and decreased mitochondrial Ca(2+) ([Ca(2+)]m) in microscopic and fluorescence‐activated cell sorting analyses, resulting in augmented eNOS Ser1177 phosphorylation and decreased eNOS Thr495 phosphorylation through Ca(2+)/Ca(2+)/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 Ca(2+) 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 [Ca(2+)]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 Ca(2+)/Ca(2+)/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 Ca(2+)‐dependent eNOS phosphorylation. This novel mechanism drives arginase activation, mitochondrial dysfunction, endothelial dysfunction, and atherogenesis.
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spelling pubmed-62229412018-11-19 Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner Koo, Bon‐Hyeock Hwang, Hye‐Mi Yi, Bong‐Gu Lim, Hyun Kyo Jeon, Byeong Hwa Hoe, Kwang Lae Kwon, Young‐Guen Won, Moo‐Ho Kim, Young Myeong Berkowitz, Dan E. Ryoo, Sungwoo J Am Heart Assoc Original Research 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 Ca(2+)/Ca(2+)/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 Ca(2+) ([Ca(2+)]c) and decreased mitochondrial Ca(2+) ([Ca(2+)]m) in microscopic and fluorescence‐activated cell sorting analyses, resulting in augmented eNOS Ser1177 phosphorylation and decreased eNOS Thr495 phosphorylation through Ca(2+)/Ca(2+)/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 Ca(2+) 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 [Ca(2+)]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 Ca(2+)/Ca(2+)/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 Ca(2+)‐dependent eNOS phosphorylation. This novel mechanism drives arginase activation, mitochondrial dysfunction, endothelial dysfunction, and atherogenesis. John Wiley and Sons Inc. 2018-09-07 /pmc/articles/PMC6222941/ /pubmed/30371203 http://dx.doi.org/10.1161/JAHA.118.009579 Text en © 2018 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Research
Koo, Bon‐Hyeock
Hwang, Hye‐Mi
Yi, Bong‐Gu
Lim, Hyun Kyo
Jeon, Byeong Hwa
Hoe, Kwang Lae
Kwon, Young‐Guen
Won, Moo‐Ho
Kim, Young Myeong
Berkowitz, Dan E.
Ryoo, Sungwoo
Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title_full Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title_fullStr Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title_full_unstemmed Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title_short Arginase II Contributes to the Ca(2+)/CaMKII/eNOS Axis by Regulating Ca(2+) Concentration Between the Cytosol and Mitochondria in a p32‐Dependent Manner
title_sort arginase ii contributes to the ca(2+)/camkii/enos axis by regulating ca(2+) concentration between the cytosol and mitochondria in a p32‐dependent manner
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6222941/
https://www.ncbi.nlm.nih.gov/pubmed/30371203
http://dx.doi.org/10.1161/JAHA.118.009579
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