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dc.contributor.authorHryciw, DH
dc.contributor.authorJenkin, KA
dc.contributor.authorSimcocks, AC
dc.contributor.authorGrinfeld, E
dc.contributor.authorMcAinch, AJ
dc.contributor.authorPoronnik, P
dc.date.accessioned2017-07-17T05:53:52Z
dc.date.available2017-07-17T05:53:52Z
dc.date.issued2012
dc.identifier.issn1357-2725
dc.identifier.doi10.1016/j.biocel.2012.02.007
dc.identifier.urihttp://hdl.handle.net/10072/341927
dc.description.abstractAlbumin endocytosis in the proximal tubule is mediated by a number of proteins, including the scavenger receptor megalin/cubilin and the PSD-95/Dlg/ZO-1 (PDZ) scaffolds NHERF1 and NHERF2. In addition, in a number of in vitro and in vivo models, the loss of ClC-5 results in a decreased cell surface expression and whole cell level of megalin, suggesting an interaction between these two proteins in vivo. We investigated if ClC-5 and megalin interact directly, and as ClC-5 binds to NHERF2, we investigated if this PDZ scaffold was required for a megalin/ClC-5 complex. GST-pulldown and immunoprecipitation experiments using rat kidney lysate demonstrated an interaction between ClC-5 and megalin, which was mediated by their C-termini. As this interaction may be controlled by a scaffold protein, we characterised any interaction between megalin and NHERF2. Immunoprecipitation experiments indicated that megalin interacts with NHERF2 in vivo, and that this interaction was via an internal NHERF binding domain in the C-terminus of megalin and PDZ2 and the C-terminus of NHERF2. Silencing NHERF2 had no effect on megalin protein levels in the whole cell or plasma membrane. Using siRNA against NHERF2, we demonstrated that NHERF2 was required to facilitate the interaction between megalin and ClC-5. Using fusion proteins, we characterised a protein complex containing ClC-5 and megalin, which is scaffolded by NHERF2, in the absence of any other proteins. Importantly, these observations are the first to describe an interaction between megalin and ClC-5, which is scaffolded by NHERF2 in proximal tubule cells.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherPergamon
dc.relation.ispartofpagefrom815
dc.relation.ispartofpageto823
dc.relation.ispartofissue5
dc.relation.ispartofjournalInternational Journal of Biochemistry and Cell Biology
dc.relation.ispartofvolume44
dc.subject.fieldofresearchBiochemistry and cell biology
dc.subject.fieldofresearchBiochemistry and cell biology not elsewhere classified
dc.subject.fieldofresearchMedical biochemistry and metabolomics
dc.subject.fieldofresearchMedical physiology
dc.subject.fieldofresearchcode3101
dc.subject.fieldofresearchcode310199
dc.subject.fieldofresearchcode3205
dc.subject.fieldofresearchcode3208
dc.titleThe interaction between megalin and ClC-5 is scaffolded by the Na+–H+ exchanger regulatory factor 2 (NHERF2) in proximal tubule cells
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.hasfulltextNo Full Text
gro.griffith.authorSkelly, Deanne


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