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Heat Shock Protein 70 and CHIP Promote Nox4 Ubiquitination and Degradation within the Losartan Antioxidative Effect in Proximal Tubule Cells.
Gil Lorenzo, Andrea F; Costantino, Valeria V; Appiolaza, Martin López; Cacciamani, Valeria; Benardon, Maria E; Bocanegra, Victoria; Vallés, Patricia G.
Afiliación
  • Gil Lorenzo AF; IMBECU-CONICET (National Council of Scientific and Technical Research of Argentina), Universidad Nacional de Cuyo, Mendoza, Argentina.
Cell Physiol Biochem ; 36(6): 2183-97, 2015.
Article en En | MEDLINE | ID: mdl-26279425
BACKGROUND: Angiotensin II/Angiotensin II type 1 receptor (AT1R) effects are dependent on ROS production stimulated by NADPH oxidase activation. Hsp70 regulates a diverse set of signaling pathways through their interactions with proteins. CHIP is a E3 ubiquitin ligase that targets proteins for polyubiquitination and degradation. AIM: We study whether Hsp70/CHIP contribute to the negative regulation of Nox4 after AT1R blockage. METHODS/RESULTS: Primary culture of proximal tubule epithelial cells (PTCs) from SHR and WKY were stimulated with Angiotensin II (AII) or treated with Losartan (L) or Losartan plus Angiotensin II (L+AII). Losartan decreased AT1R and Nox4 while enhancing caveolin-1 and Hsp70 protein expression in SHR PTCs. Immunoprecipitation and immunofluorescence proved interaction and colocalization of increased Hsp70/CHIP with decreased Nox4 in SHR PTCs (L) vs (All). Hsp72 knockdown resulted in enhanced Nox4 protein levels, NADPH oxidase activity and ROS generation in (L+AII) revealing that Losartan was unable to abrogate AII effects on Nox4 expression and oxidative activity. Moreover, MG132 exposed PTCs (L) demostrated blocked ubiquitinated Nox4 degradation and increased colocalization of Nox4/Ubiquitin by inmunofluorescence. Conversely, Hsp72 depletion reduced Nox4/Ubiquitin colocalization causing Nox4 upregulation due to proteosomal degradation inhibition, although Losartan treatment. CONCLUSION: Our study demonstrates that Hsp70 and CHIP mediates the ubiquitination and proteasomal degradation of Nox4 as part of the antioxidative effect of Losartan in SHR.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas HSP70 de Choque Térmico / NADPH Oxidasas / Losartán / Ubiquitina-Proteína Ligasas / Ubiquitinación / Proteolisis / Túbulos Renales Proximales / Antioxidantes Límite: Animals / Humans / Male Idioma: En Revista: Cell Physiol Biochem Asunto de la revista: BIOQUIMICA / FARMACOLOGIA Año: 2015 Tipo del documento: Article País de afiliación: Argentina Pais de publicación: Alemania

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas HSP70 de Choque Térmico / NADPH Oxidasas / Losartán / Ubiquitina-Proteína Ligasas / Ubiquitinación / Proteolisis / Túbulos Renales Proximales / Antioxidantes Límite: Animals / Humans / Male Idioma: En Revista: Cell Physiol Biochem Asunto de la revista: BIOQUIMICA / FARMACOLOGIA Año: 2015 Tipo del documento: Article País de afiliación: Argentina Pais de publicación: Alemania