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J Physiol ; 551(Pt 2): 455-66, 2003 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-12816971

RESUMEN

The serum- and glucocorticoid-induced kinase-1 (sgk1) increases the activity of a number of epithelial ion channels and transporters. The present study examines the distribution and subcellular localization of sgk1 protein in the rat kidney and the regulation of levels of expression induced by steroids. The results indicate that the kidney expresses predominantly the sgk1 isoform with a distribution restricted to the thick ascending limb of Henle, distal convoluted, connecting and cortical collecting tubules. Within cells, sgk1 strongly associates with the microsomal fraction of homogenates and it colocalizes with the Na+,K+-ATPase to the basolateral membrane. Analysis of the levels of expression of sgk1 by Western blotting and immunohistochemistry indicates constitutive high expression under basal conditions. Approximately half of the basal level is maintained by glucocorticoids whereas physiological fluctuations of aldosterone produce minor changes in sgk1 abundance in adrenal-intact animals. These results do not support the notion that physiological changes of aldosterone concentration turn the expression of sgk1 'on and off' in the mammalian kidney. Additionally, localization of sgk1 to the basolateral membrane indicates that the effects mediated by sgk1 do not require a direct interaction with the ion channels and transporters whose activity is modulated, since most of these proteins are located in the apical membrane of renal epithelial cells.


Asunto(s)
Regulación Enzimológica de la Expresión Génica/fisiología , Riñón/enzimología , Proteínas Nucleares , Proteínas Serina-Treonina Quinasas/metabolismo , Adrenalectomía , Aldosterona/farmacología , Animales , Anticuerpos/química , Especificidad de Anticuerpos , Northern Blotting , Western Blotting , Células Cultivadas , ADN Complementario/biosíntesis , ADN Complementario/genética , Electroforesis en Gel de Poliacrilamida , Células Epiteliales/enzimología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Glucocorticoides/farmacología , Proteínas Inmediatas-Precoces , Immunoblotting , Técnicas In Vitro , Isoenzimas/genética , Isoenzimas/inmunología , Isoenzimas/metabolismo , Túbulos Renales/enzimología , Microscopía Fluorescente , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/inmunología , ARN/aislamiento & purificación , ARN/metabolismo , Ratas , Ratas Sprague-Dawley , Fracciones Subcelulares/enzimología , Transfección
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