Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture
Angiotensin converting enzyme 2 (ACE2) is a component of the renin-angiotensin system (RAS) which converts Ang II, a potent vasoconstrictor peptide into Ang 1-7, a vasodilator peptide which may act as a negative feedback hormone to the actions of Ang II. the discovery of this enzyme added a new leve...
| Autores: | , , , , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2011 |
| País: | Brasil |
| Institución: | Universidade Federal de São Paulo (UNIFESP) |
| Repositorio: | Repositório Institucional da UNIFESP |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.unifesp.br:11600/33814 |
| Acceso en línea: | http://dx.doi.org/10.1016/j.ijbiomac.2011.03.018 http://repositorio.unifesp.br/handle/11600/33814 |
| Access Level: | acceso abierto |
| Palabra clave: | ACE2 Mesangial cells Renin-angiotensin system |
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Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in cultureACE2Mesangial cellsRenin-angiotensin systemAngiotensin converting enzyme 2 (ACE2) is a component of the renin-angiotensin system (RAS) which converts Ang II, a potent vasoconstrictor peptide into Ang 1-7, a vasodilator peptide which may act as a negative feedback hormone to the actions of Ang II. the discovery of this enzyme added a new level of complexity to this system. the mesangial cells (MC) have multiple functions in glomerular physiology and pathophysiology and are able to express all components of the RAS. Despite of being localized in these cells, ACE2 has not yet been purified or characterized. in this study ACE2 from mice immortalized MC (IMC) was purified by ion-exchange chromatography. the purified enzyme was identified as a single band around 60-70 kDa on SDS-polyacrylamide gel and by Western blotting using a specific antibody. the optima pH and chloride concentrations were 7.5 and 200 mM, respectively. the N-terminal sequence was homologous with many species ACE2 N-terminal sequences as described in the literature. ACE2 purified from IMC was able to hydrolyze Ang II into Ang 1-7 and the K(m) value for Ang II was determined to be 2.87 +/- 0.76 mu M. in conclusion, we purified and localized, for the first time, ACE2 in MC, which was able to generate Ang 1-7 from Ang II. Ang 1-7 production associated to Ang II degradation by ACE2 may exert a protective effect in the renal hemodynamic. (C) 2011 Elsevier B.V. All rights reserved.Universidade Federal de São Paulo, Div Nephrol, Dept Med, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Immunol & Parasitol Div, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Div Nephrol, Dept Med, BR-04023900 São Paulo, BrazilUniversidade Federal de São Paulo, Immunol & Parasitol Div, BR-04023900 São Paulo, BrazilWeb of ScienceFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 05-57543-0Elsevier B.V.Universidade Federal de São Paulo (UNIFESP)2016-01-24T14:16:54Z2016-01-24T14:16:54Z2011-07-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion79-84application/pdfhttp://dx.doi.org/10.1016/j.ijbiomac.2011.03.018International Journal of Biological Macromolecules. Amsterdam: Elsevier B.V., v. 49, n. 1, p. 79-84, 2011.10.1016/j.ijbiomac.2011.03.018WOS000291840400012.pdf0141-8130http://repositorio.unifesp.br/handle/11600/33814WOS:000291840400012ark:/48912/001300002n9hjengInternational Journal of Biological Macromoleculesinfo:eu-repo/semantics/openAccesshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPAragao, Danielle Sanches [UNIFESP]Cunha, Tatiana de Sousa [UNIFESP]Arita, Danielle Yuri [UNIFESP]Andrade, Maria Claudina Camargo de [UNIFESP]Fernandes, Adriana Barrinha [UNIFESP]Watanabe, Ingrid Kazue Mizuno [UNIFESP]Mortara, Renato Arruda [UNIFESP]Casarini, Dulce Elena [UNIFESP]2024-07-31T20:43:36Zoai:repositorio.unifesp.br:11600/33814Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-07-31T20:43:36Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
| dc.title.none.fl_str_mv |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| title |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| spellingShingle |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture Aragao, Danielle Sanches [UNIFESP] ACE2 Mesangial cells Renin-angiotensin system |
| title_short |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| title_full |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| title_fullStr |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| title_full_unstemmed |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| title_sort |
Purification and characterization of angiotensin converting enzyme 2 (ACE2) from murine model of mesangial cell in culture |
| dc.creator.none.fl_str_mv |
Aragao, Danielle Sanches [UNIFESP] Cunha, Tatiana de Sousa [UNIFESP] Arita, Danielle Yuri [UNIFESP] Andrade, Maria Claudina Camargo de [UNIFESP] Fernandes, Adriana Barrinha [UNIFESP] Watanabe, Ingrid Kazue Mizuno [UNIFESP] Mortara, Renato Arruda [UNIFESP] Casarini, Dulce Elena [UNIFESP] |
| author |
Aragao, Danielle Sanches [UNIFESP] |
| author_facet |
Aragao, Danielle Sanches [UNIFESP] Cunha, Tatiana de Sousa [UNIFESP] Arita, Danielle Yuri [UNIFESP] Andrade, Maria Claudina Camargo de [UNIFESP] Fernandes, Adriana Barrinha [UNIFESP] Watanabe, Ingrid Kazue Mizuno [UNIFESP] Mortara, Renato Arruda [UNIFESP] Casarini, Dulce Elena [UNIFESP] |
| author_role |
author |
| author2 |
Cunha, Tatiana de Sousa [UNIFESP] Arita, Danielle Yuri [UNIFESP] Andrade, Maria Claudina Camargo de [UNIFESP] Fernandes, Adriana Barrinha [UNIFESP] Watanabe, Ingrid Kazue Mizuno [UNIFESP] Mortara, Renato Arruda [UNIFESP] Casarini, Dulce Elena [UNIFESP] |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) |
| dc.subject.por.fl_str_mv |
ACE2 Mesangial cells Renin-angiotensin system |
| topic |
ACE2 Mesangial cells Renin-angiotensin system |
| description |
Angiotensin converting enzyme 2 (ACE2) is a component of the renin-angiotensin system (RAS) which converts Ang II, a potent vasoconstrictor peptide into Ang 1-7, a vasodilator peptide which may act as a negative feedback hormone to the actions of Ang II. the discovery of this enzyme added a new level of complexity to this system. the mesangial cells (MC) have multiple functions in glomerular physiology and pathophysiology and are able to express all components of the RAS. Despite of being localized in these cells, ACE2 has not yet been purified or characterized. in this study ACE2 from mice immortalized MC (IMC) was purified by ion-exchange chromatography. the purified enzyme was identified as a single band around 60-70 kDa on SDS-polyacrylamide gel and by Western blotting using a specific antibody. the optima pH and chloride concentrations were 7.5 and 200 mM, respectively. the N-terminal sequence was homologous with many species ACE2 N-terminal sequences as described in the literature. ACE2 purified from IMC was able to hydrolyze Ang II into Ang 1-7 and the K(m) value for Ang II was determined to be 2.87 +/- 0.76 mu M. in conclusion, we purified and localized, for the first time, ACE2 in MC, which was able to generate Ang 1-7 from Ang II. Ang 1-7 production associated to Ang II degradation by ACE2 may exert a protective effect in the renal hemodynamic. (C) 2011 Elsevier B.V. All rights reserved. |
| publishDate |
2011 |
| dc.date.none.fl_str_mv |
2011-07-01 2016-01-24T14:16:54Z 2016-01-24T14:16:54Z |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.ijbiomac.2011.03.018 International Journal of Biological Macromolecules. Amsterdam: Elsevier B.V., v. 49, n. 1, p. 79-84, 2011. 10.1016/j.ijbiomac.2011.03.018 WOS000291840400012.pdf 0141-8130 http://repositorio.unifesp.br/handle/11600/33814 WOS:000291840400012 |
| dc.identifier.dark.fl_str_mv |
ark:/48912/001300002n9hj |
| url |
http://dx.doi.org/10.1016/j.ijbiomac.2011.03.018 http://repositorio.unifesp.br/handle/11600/33814 |
| identifier_str_mv |
International Journal of Biological Macromolecules. Amsterdam: Elsevier B.V., v. 49, n. 1, p. 79-84, 2011. 10.1016/j.ijbiomac.2011.03.018 WOS000291840400012.pdf 0141-8130 WOS:000291840400012 ark:/48912/001300002n9hj |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
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International Journal of Biological Macromolecules |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
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openAccess |
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http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
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79-84 application/pdf |
| dc.publisher.none.fl_str_mv |
Elsevier B.V. |
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Elsevier B.V. |
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reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
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Universidade Federal de São Paulo (UNIFESP) |
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UNIFESP |
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UNIFESP |
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Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
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