Nedd4-2 functionally interacts with ClC-5: Involvement in constitutive albumin endocytosis in proximal tubule cells

Deanne H. Hryciw; Jenny Ekberg; Aven Lee; Ingrid L. Lensink; Sharad Kumar; William B. Guggino; David I. Cook; Carol A. Pollock; Philip Poronnik

doi:10.1074/jbc.M411491200

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Nedd4-2 functionally interacts with ClC-5: Involvement in constitutive albumin endocytosis in proximal tubule cells

Journal article

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Nedd4-2 functionally interacts with ClC-5: Involvement in constitutive albumin endocytosis in proximal tubule cells

Deanne H. Hryciw, Jenny Ekberg, Aven Lee, Ingrid L. Lensink, Sharad Kumar, William B. Guggino, David I. Cook, Carol A. Pollock and Philip Poronnik

Journal of Biological Chemistry, Vol.279(53), pp.54996-55007

2004

DOI: https://doi.org/10.1074/jbc.M411491200

PMID: 15489223

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Abstract

Membrane Transport

Structure

Function

Biogenesis

Constitutive albumin uptake by the proximal tubule is achieved by a receptor-mediated process in which the Cl(-) channel, ClC-5, plays an obligate role. Here we investigated the functional interaction between ClC-5 and ubiquitin ligases Nedd4 and Nedd4-2 and their role in albumin uptake in opossum kidney proximal tubule (OK) cells. In vivo immunoprecipitation using an anti-HECT antibody demonstrated that ClC-5 bound to ubiquitin ligases, whereas glutathione S-transferase pull-downs confirmed that the C terminus of ClC-5 bound both Nedd4 and Nedd4-2. Nedd4-2 alone was able to alter ClC-5 currents in Xenopus oocytes by decreasing cell surface expression of ClC-5. In OK cells, a physiological concentration of albumin (10 mug/ml) rapidly increased cell surface expression of ClC-5, which was also accompanied by the ubiquitination of ClC-5. Albumin uptake was reduced by inhibiting either the lysosome or proteasome. Total levels of Nedd4-2 and proteasome activity also increased rapidly in response to albumin. Overexpression of ligase defective Nedd4-2 or knockdown of endogenous Nedd4-2 with small interfering RNA resulted in significant decreases in albumin uptake. In contrast, pathophysiological concentrations of albumin (100 and 1000 mug/ml) reduced the levels of ClC-5 and Nedd4-2 and the activity of the proteasome to the levels seen in the absence of albumin. These data demonstrate that normal constitutive uptake of albumin by the proximal tubule requires Nedd4-2, which may act via ubiquitination to shunt ClC-5 into the endocytic pathway.

Details

Title: Nedd4-2 functionally interacts with ClC-5: Involvement in constitutive albumin endocytosis in proximal tubule cells
Authors: Deanne H. Hryciw - The University of Queensland
Jenny Ekberg - The University of Queensland
Aven Lee - The University of Queensland
Ingrid L. Lensink - Hanson Institute
Sharad Kumar - Hanson Institute
William B. Guggino - Johns Hopkins University
David I. Cook - The University of Sydney
Carol A. Pollock - The University of Sydney
Philip Poronnik - The University of Queensland
Publication details: Journal of Biological Chemistry, Vol.279(53), pp.54996-55007
Publisher: Elsevier Inc.
Date published: 2004
DOI: 10.1074/jbc.M411491200
ISSN: 1083-351X; 0021-9258
PMID: 15489223
Grant note: This work was funded by grants from the NHRMC and Juvenile Diabetes Fund International (to P. P. and C. A. P.) and National Institutes of Health Grants DK32753 and HL47122 (to W. B. G.).
Organisation Unit: School of Health - Biomedicine
Language: English
Record Identifier: 991186523502621
Output Type: Journal article

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