Abstract

The renal Na+-K+-2Cl- co-transporter (NKCC2) is expressed in kidney thick ascending limb (TAL) cells, where it mediates NaCl reabsorption regulating body salt levels and blood pressure. In this study we used a well-characterized NKCC2 construct (c-NKCC2) to identify NKCC2 interacting protein by an antibody shift assay coupled with Blue Native/SDS-PAGE (BN/SDS-PAGE) and Mass Spectrometry (MS). Among the interacting proteins we identified moesin, a protein belonging to ERM (Ezrin, Radixin, Moesin) family. Co-immunoprecipitation experiments confirmed that c-NKCC2 interacts with the N-terminal domain of moesin in LLC-PK1 cells. Moreover, c-NKCC2 accumulates in intracellular and sub-apical vesicles in cells transfected with a moesin dominant negative GFP-tagged construct. In addition, moesin knockdown by siRNA decreases by about 50% c-NKCC2 surface expression. Specifically, endocytosis and exocytosis assays showed that moesin knockdown does not affect NKCC2 internalization but strongly reduces exocytosis of the co-transporter. Our data clearly demonstrate that moesin plays a critical role in apical membrane insertion of NKCC2, suggesting a possible involvement of moesin in regulation of Na+ and Cl- absorption in the kidney.

Autore Pugliese

• Valenti Giovanna , Procino Giuseppe

Tutti gli autori

• VALENTI G.;PROCINO G.;SVELTO M.

Titolo volume/Rivista

Non Disponibile

Anno di pubblicazione

2011

ISSN

1748-1708

ISBN

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Numero di citazioni Wos

Nessuna citazione

Ultimo Aggiornamento Citazioni

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Numero di citazioni Scopus

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Ultimo Aggiornamento Citazioni

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Settori ERC

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Codici ASJC

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