Wiley Online Library User AgreementMathilde KeckMathieu GeneteJacques TeulonStéphane LourdelKamel LaghmaniMathilde KeckThomas PennaforteThomas PennaforteMathieu GeneteJacques TeulonTeddy GrandTeddy GrandSébastien L'HosteSébastien L'HosteNadia DefontaineNadia DefontaineStéphane LourdelKamel LaghmaniDavid MordasiniDavid Mordasini2025-06-182025-06-182011-03-151059-77941098-100410.1002/humu.21467https://trapdev.rcub.bg.ac.rs/handle/123456789/1035797Mutations in the electrogenic Cl(-)/H(+) exchanger ClC-5 gene CLCN5 are frequently associated with Dent disease, an X-linked recessive disorder affecting the proximal tubules. Here, we investigate the consequences in Xenopus laevis oocytes and in HEK293 cells of nine previously reported, pathogenic, missense mutations of ClC-5, most of them which are located in regions forming the subunit interface. Two mutants trafficked normally to the cell surface and to early endosomes, and displayed complex glycosylation at the cell surface like wild-type ClC-5, but exhibited reduced currents. Three mutants displayed improper N-glycosylation, and were nonfunctional due to being retained and degraded at the endoplasmic reticulum. Functional characterization of four mutants allowed us to identify a novel mechanism leading to ClC-5 dysfunction in Dent disease. We report that these mutant proteins were delayed in their processing, and that the stability of their complex glycosylated form was reduced, causing lower cell surface expression. The early endosome distribution of these mutants was normal. Half of these mutants displayed reduced currents, whereas the other half showed abolished currents. Our study revealed distinct cellular mechanisms accounting for ClC-5 loss of function in Dent disease.OPENERL 7226 KeckCNRS[SDV]Life Sciences [q-bio]NadiaMolecular Sequence DataERL 7226 LourdelChloride/proton exchangerClC-5Kamel[SDV.BC]Life Sciences [q-bio]/Cellular BiologyProcessingERL 7226 L'HosteMathildeKidney Tubules, ProximalXenopus laevisChloride ChannelsJacquesStéphaneAnimalsHumansAmino Acid SequenceSébastienCells, CulturedUPMC Univ Paris 06INSERMERL 7226 PennaforteDent DiseaseTeddyMathieuLife SciencesERL 7226 TeulonComplete List of Authors: GrandDavidThomasHEK293 CellsERL 7226 Key Words: Dent's diseaseMutationOocytesERL 7226 DefontaineCLCN5UMR_S 872ERL 7226 MordasiniSequence AlignmentERL 7226 LaghmaniERL 7226 Genetepublication0301 basic medicine03 medical and health sciencesdoi_dedup___:b2c610cab3ecb264829d09f56c8e231021305656