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- W2015215948 abstract "The chloride/proton transporter ClC-5 mediates the re-absorption of filtered proteins in the kidney by promoting the formation of the macromolecular endocytic complex and by aiding in endosomal acidification upon complex internalization in proximal tubule cells. Mutations disrupting ClC-5 lead to proteinuria reflecting a severe impairment of renal receptor endocytosis and several disease-causing mutations in ClC-5 translate into truncations of the carboxy terminus (Ct), highlighting the significance of this region. The Ct interacts with the scaffold protein NHERF2 (Na+/H+ exchanger regulatory factor 2) and leads to enhanced endocytic uptake of albumin by kidney cells [Hryciw, D. et al. 2006]. This interaction occurs through the second PDZ domain of NHERF2 and an unknown binding site on the Ct of ClC-5 [Hryciw, D. et al. 2006]. In the present study NHERF2/ClC-5 Ct interactions were confirmed and a putative internal PDZ binding motif TSII (residues 657-60) was identified in the Ct. Mapping of this motif on the crystal structure of ClC-5 reveals it lies up-stream of a β-turn, a secondary structure element thought to be critical for PDZ domain recognition of an internal motif [Hillier, B. et al. 1999]. The strategic S658A mutation completely abolishes NHERF2 binding in vitro suggesting the PDZ binding motif was targeted. CD analysis confirmed similar spectra between wildtype and S658A mutant, indicating that this mutation does not lead to gross misfolding of the Ct. The disease mutant R648X which removes this critical S658 abolishes NHERF2 interactions providing further evidence the internal PDZ binding motif was targeted. Future work will focus on identifying proteins interacting with the ClC-5/NHERF2 complex and deciphering the role of this complex in renal endocytosis. This work was supported by the Kidney Foundation of Canada operating gant to C.B. and a NSERC studentship to L.W." @default.
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- W2015215948 date "2009-02-01" @default.
- W2015215948 modified "2023-09-28" @default.
- W2015215948 title "Identification Of The NHERF2 Binding Site For The Chloride/Proton Transporter ClC-5" @default.
- W2015215948 doi "https://doi.org/10.1016/j.bpj.2008.12.2289" @default.
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