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dc.contributor.authorHryciw, DH
dc.contributor.authorEkberg, J
dc.contributor.authorLee, A
dc.contributor.authorLensink, IL
dc.contributor.authorKumar, S
dc.contributor.authorGuggino, WB
dc.contributor.authorCook, DI
dc.contributor.authorPollock, CA
dc.contributor.authorPoronnik, P
dc.date.accessioned2017-05-03T16:58:10Z
dc.date.available2017-05-03T16:58:10Z
dc.date.issued2004
dc.date.modified2010-01-07T02:17:50Z
dc.identifier.issn0021-9258
dc.identifier.doi10.1074/jbc.M411491200
dc.identifier.urihttp://hdl.handle.net/10072/28097
dc.description.abstractConstitutive 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 姯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 姯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.
dc.description.peerreviewedYes
dc.description.publicationstatusYes
dc.languageEnglish
dc.language.isoeng
dc.publisherThe American Society for Biochemistry and Molecular Biology
dc.publisher.placeUnited States of America
dc.relation.ispartofpagefrom54996
dc.relation.ispartofpageto55007
dc.relation.ispartofissue53
dc.relation.ispartofjournalThe Journal of Biological Chemistry
dc.relation.ispartofvolume279
dc.subject.fieldofresearchChemical sciences
dc.subject.fieldofresearchBiological sciences
dc.subject.fieldofresearchBiomedical and clinical sciences
dc.subject.fieldofresearchCellular nervous system
dc.subject.fieldofresearchcode34
dc.subject.fieldofresearchcode31
dc.subject.fieldofresearchcode32
dc.subject.fieldofresearchcode320902
dc.titleNedd4-2 Functionally Interacts with ClC-5: Involvement in Constitutive Albumin Endocytosis in Proximal Tubule Cells
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.date.issued2004
gro.hasfulltextNo Full Text
gro.griffith.authorEkberg, Jenny A.
gro.griffith.authorSkelly, Deanne


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