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Showing CLCN3 — CLC-3 is a alias.

CLCN3

H(+)/Cl(-) exchange transporter 3 · UniProt P51790

Length
818 aa
Mass
91.0 kDa
Annotated
2026-06-09
100 papers in source corpus 38 papers cited in narrative 38 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 7/7 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

CLCN3 (ClC-3) is an intracellular Cl-/H+ antiporter that provides the counter-ion conductance neutralizing the charge built up by the vacuolar H+-ATPase, thereby driving acidification of endosomal, lysosomal, and synaptic vesicle compartments (PMID:11997263, PMID:12059962, PMID:17977943). Ion coupling is governed by an extracellular gating glutamate (E224) and an internal anion-gate tyrosine (Y572) that together maintain Cl-/H+ stoichiometry through the transport cycle; mutation of E224 eliminates proton coupling and abolishes acidification, while the E224–Y572 gate also confers pronounced voltage-dependent capacitive behavior and uncoupling at low luminal pH (PMID:17977943, PMID:19926787, PMID:29917234, PMID:23509947). Three splice variants share identical outwardly rectifying transport activity but use distinct N-terminal sorting motifs to target late endosomes/lysosomes versus recycling endosomes, and the ClC-3c isoform forms heterodimers that chaperone ClC-4 out of the ER into endosomes (PMID:26342074, PMID:28972156). In vivo, ClC-3 loss raises endosomal pH and causes neurodegeneration resembling neuronal ceroid lipofuscinosis (PMID:12059962), and it is required for synaptic vesicle acidification underlying GABA loading at inhibitory terminals while limiting glutamate loading and release probability at excitatory synapses (PMID:21378974, PMID:24904288). Compartment acidification by ClC-3 further supports Nox1/Nox2 NADPH-oxidase ROS production and downstream NF-κB and ICl,swell signaling in vascular and immune cells, and is needed for insulin granule priming and exocytosis (PMID:17673675, PMID:16522634, PMID:20479003, PMID:19808023). A separate plasma-membrane pool is rapidly internalized through clathrin binding to an N-terminal dileucine cluster (PMID:17652080), and at the cell surface ClC-3 mediates Cl- currents activated by CaMKII phosphorylation at Ser109 and gated as a volume sensor through Ser51, with ROCK2 phosphorylation at Thr532 driving angiotensin II-induced currents; these activities power premitotic cytoplasmic condensation, glioma invasion and chemotaxis, and vascular smooth muscle migration (PMID:14754994, PMID:11274166, PMID:26562480, PMID:18784301, PMID:20139089, PMID:23345219). De novo heterozygous gain-of-function CLCN3 missense variants cause neurodevelopmental disorders while homozygous loss-of-function variants produce severe neurodegeneration mirroring the knockout mouse (PMID:34186028).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 1999 High

    Established that ClC-3 activity is volume-sensitive and controlled by phosphorylation, identifying an N-terminal regulatory serine as a volume sensor.

    Evidence Ser51 site-directed mutagenesis with patch clamp in cardiac and NIH/3T3 cells

    PMID:9874688

    Open questions at the time
    • Did not resolve whether the volume-activated current is carried directly by ClC-3 protein versus an associated channel
    • Endogenous kinase acting on Ser51 in situ not defined
  2. 2001 High

    Demonstrated that the CLCN3 gene encodes the ion conduction pathway itself and that CaMKII phosphorylation strongly activates its plasma-membrane current.

    Evidence Pore mutation (G280E) altering anion selectivity plus surface biotinylation and patch clamp in tsA cells

    PMID:11274166

    Open questions at the time
    • Did not identify the CaMKII phosphosite
    • Relationship of surface current to predominant intracellular pool unresolved
  3. 2002 High

    Defined ClC-3 as a provider of Cl- conductance for V-ATPase-driven vesicular acidification in vitro and showed its loss disrupts endosomal/lysosomal acidification and causes neurodegeneration in vivo.

    Evidence E224A mutagenesis with acidification assays in cell lines, and Clcn3-/- mice with endosomal pH measurement and histology

    PMID:11997263 PMID:12059962

    Open questions at the time
    • Did not yet establish electrogenic Cl-/H+ exchange versus pure channel mechanism
    • Cell-type basis of selective neurodegeneration not explained
  4. 2004 High

    Identified trafficking and gating determinants: AP-3 routes ClC-3 to synaptic vesicles, and CaMKII phosphorylates Ser109 to gate the surface current.

    Evidence AP-3-deficient mocha mice with fractionation and zinc transport assay; in vitro CaMKII phosphorylation and S109A mutagenesis with patch clamp

    PMID:14754994 PMID:15073168

    Open questions at the time
    • Relationship between Ser51 and Ser109 phosphorylation events unresolved
    • Whether AP-3 cargo function requires transport activity untested
  5. 2006 High

    Connected ClC-3-dependent endosomal acidification to NADPH oxidase signaling, showing it is required for Nox1/Nox2 ROS production in vascular smooth muscle and neutrophils.

    Evidence Reciprocal Co-IP with Nox1/p22phox plus knockdown and NF-κB assays in SMCs; Clcn3-/- neutrophil phagocytosis and oxidase assays

    PMID:16522634 PMID:17673675

    Open questions at the time
    • Whether charge neutralization is sufficient or additional scaffolding role exists not separated
    • Direct Cl- flux measurement at signaling endosomes lacking
  6. 2007 High

    Resolved the transport mechanism as electrogenic Cl-/H+ antiport gated by E224 and defined clathrin-mediated surface internalization and actin attachment.

    Evidence E224A coupling analysis by patch clamp in HEK293T; dileucine cluster mutagenesis with clathrin pulldown and pulse-chase; F-actin co-sedimentation with peptide dialysis

    PMID:17442672 PMID:17652080 PMID:17977943

    Open questions at the time
    • Structural basis of the gate not defined at this stage
    • Physiological significance of the transient surface pool versus endosomal pool unclear
  7. 2008 High

    Linked ClC-3 surface Cl- efflux to cell-volume reduction in mitosis and identified Ins(3,4,5,6)P4 as an endogenous inhibitor acting on both surface and endosomal pools.

    Evidence shRNA knockdown with premitotic condensation imaging and patch clamp; cell-permeant Ins(3,4,5,6)P4 with endosomal FITC-transferrin pH imaging and neuronal recordings

    PMID:18784301 PMID:18923035 PMID:18951024

    Open questions at the time
    • Mechanism by which Ins(3,4,5,6)P4 acts on the transporter unknown
    • Direct versus indirect role in dense-core vesicle exocytosis not separated
  8. 2009 High

    Established a metabolic role: ClC-3 is required for insulin granule proton transport, priming, and depolarization-evoked exocytosis, and refined the pH-dependence of its coupling.

    Evidence Clcn3-/- beta cell capacitance and granule proton transport assays; patch clamp pH titration with E224A and cysteine mutants

    PMID:19808023 PMID:19926787

    Open questions at the time
    • Whether granule acidification defect fully accounts for priming deficit untested
    • Physiological role of low-pH uncoupling unclear
  9. 2010 High

    Demonstrated CaMKII directly binds ClC-3 to drive glioma invasion and placed endosomal ClC-3/Nox1 ROS upstream of TNF-α-activated swelling Cl- current.

    Evidence Co-IP in glioblastoma tissue with shRNA and invasion assay; Clcn3-/- VSMC patch clamp with dominant-negative Rab5/Rab11 epistasis

    PMID:20139089 PMID:20479003

    Open questions at the time
    • How endosomal ROS feeds back to activate a Cl- conductance mechanistically undefined
    • Identity of the swelling-activated current carrier not fully resolved
  10. 2011 High

    Defined ClC-3 as required for inhibitory synaptic vesicle acidification and GABA loading, and CaMKII-dependent activation drives mitotic Cl- efflux.

    Evidence Clcn3-/- hippocampal slice mIPSC recordings with VGAT colocalization and vesicle acidification assay; glioma PMC imaging with KN-93 and shRNA

    PMID:21378974 PMID:22049206

    Open questions at the time
    • Splice variant responsible for vesicular localization not assigned here
    • Quantitative contribution of Cl- versus H+ to neurotransmitter loading unresolved
  11. 2012 High

    Established opposing synaptic roles of ClC-3 at excitatory terminals and postsynaptic sites: limiting glutamate loading/release while moderating NMDA-dependent LTP via CaMKII phosphorylation.

    Evidence Clcn3-/- hippocampal mEPSC recordings and EM; LTP recordings with CaMKII-site decoy peptide and NMDA receptor Co-IP

    PMID:17046694 PMID:23165767 PMID:24904288

    Open questions at the time
    • How a vesicular transporter and a postsynaptic surface channel pool are partitioned in the same neuron unclear
    • Cell-cycle-dependent nucleocytoplasmic localization (#37) mechanistically unexplained
  12. 2013 High

    Characterized ClC-3's specialized capacitive non-transporting cycles and showed CaMKII activation links bradykinin signaling to glioma chemotaxis.

    Evidence Electrophysiology with fluorescence pH and proton-glutamate mutagenesis using an N-terminal retention mutant; simultaneous Ca2+ imaging/patch clamp with shRNA and chemotaxis assay

    PMID:23345219 PMID:23509947

    Open questions at the time
    • Functional significance of capacitive cycles in native compartments unknown
    • Direct demonstration that surface ClC-3 carries the bradykinin-evoked current limited
  13. 2016 High

    Identified ROCK2 phosphorylation at Thr532 and Endophilin A2 SH3 binding as regulators of ClC-3 surface trafficking and angiotensin/volume-regulated currents in vascular smooth muscle.

    Evidence T532D/T532A mutagenesis with reciprocal Co-IP, ROCK2 siRNA, patch clamp and migration assay; Endophilin A2 Co-IP and live imaging in transgenic mice

    PMID:26562480 PMID:27760895

    Open questions at the time
    • How three phosphosites (Ser51, Ser109, Thr532) are coordinated unresolved
    • Endophilin A2 interaction confirmed in a single lab
  14. 2017 High

    Showed the ClC-3c isoform chaperones ClC-4 from ER to endosomes via heterodimer formation, defining an assembly partnership among CLC family members.

    Evidence Clear-native gel electrophoresis and localization in Clcn3-/- astrocytes with co-expression in HEK293T

    PMID:28972156

    Open questions at the time
    • Functional consequence of heterodimeric versus homodimeric transport not quantified
    • Whether heterodimerization alters ion coupling untested
  15. 2018 High

    Mapped the E224–Y572 gate as the structural determinant of Cl-/H+ coupling and gating charge.

    Evidence Systematic mutagenesis (E224A, Y572S/F, M531A) with biophysical coupling and gating-charge analysis by patch clamp

    PMID:29917234

    Open questions at the time
    • Atomic structure of the gate not determined
    • How regulatory phosphorylation alters gate behavior unknown
  16. 2021 High

    Established CLCN3 as a human disease gene, linking gain-of-function variants to neurodevelopmental disorders and loss-of-function variants to severe neurodegeneration.

    Evidence Electrophysiology of patient variants in oocytes and mammalian cells with clinical/MRI phenotyping

    PMID:34186028

    Open questions at the time
    • Cellular consequence of altered luminal pH inhibition in patient neurons not directly shown
    • Genotype-phenotype relationship across the variant spectrum incomplete

Open questions

Synthesis pass · forward-looking unresolved questions
  • How the single ClC-3 protein partitions among its intracellular transporter, surface CaMKII-gated channel, and capacitive roles, and how multiple phosphorylation and lipid signals are integrated, remains unresolved.
  • No high-resolution structure linking gating residues to regulatory sites
  • Quantitative balance between intracellular and plasma-membrane pools in native tissue undefined
  • Mechanism coupling endosomal ROS production to surface Cl- currents unclear

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005215 transporter activity 5 GO:0140098 catalytic activity, acting on RNA 3 GO:0008092 cytoskeletal protein binding 1 GO:0098772 molecular function regulator activity 1
Localization
GO:0005764 lysosome 4 GO:0005768 endosome 4 GO:0005886 plasma membrane 4 GO:0031410 cytoplasmic vesicle 4 GO:0005783 endoplasmic reticulum 1
Pathway
R-HSA-112316 Neuronal System 4 R-HSA-9609507 Protein localization 4 R-HSA-1852241 Organelle biogenesis and maintenance 3 R-HSA-8953897 Cellular responses to stimuli 3 R-HSA-1640170 Cell Cycle 2
Partners
CAMK2CLCN4EBP50GRIN1NOX1ROCK2SH3GL1VDAC1
Complex memberships
ClC-3/ClC-4 heterodimer

Evidence

Reading pass · 38 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1999 A serine residue at position 51 (Ser51) in the intracellular N-terminus of ClC-3, within a PKC consensus phosphorylation site, acts as a key volume sensor. PKC phosphorylation closes the channel, while inhibition of PKC or cell swelling opens it. Site-directed mutagenesis of Ser51 abolished volume-regulated ClC-3 channel activity. Site-directed mutagenesis combined with patch-clamp electrophysiology in cardiac cells and NIH/3T3 cells The Journal of general physiology High 9874688
2001 CaMKII regulates ClC-3 channel activity by phosphorylation, increasing plasma membrane Cl- current ~22-fold. A G280E mutation in the pore region changed anion selectivity from I->Cl- to Cl->I-, confirming ClC-3 encodes the channel. Surface biotinylation demonstrated plasma membrane expression. Whole-cell patch clamp, surface biotinylation, site-directed mutagenesis in tsA cells stably transfected with hCLC-3 The Journal of biological chemistry High 11274166
2002 Heterologous ClC-3 expression promotes lysosomal acidification by providing Cl- conductance for charge neutralization of the vacuolar H+-ATPase. An E224A channel-activity mutant failed to induce vesicle formation, demonstrating that Cl- channel activity is required for acidification. Transfection in CHO-K1 and Huh-7 cells, bafilomycin inhibition, colocalization with LAMP-1/LAMP-2/cathepsin D, E224A mutagenesis American journal of physiology. Cell physiology High 11997263
2002 ClC-3 deficiency in mice causes elevated endosomal pH and progressive neurodegeneration resembling neuronal ceroid lipofuscinosis (NCL), with lysosomal accumulation of mitochondrial F1F0-ATPase subunit c, demonstrating ClC-3 is required for normal endosomal/lysosomal acidification in vivo. Targeted gene disruption (Clcn3-/- mice), immunohistochemistry, Western blot, endosomal pH measurement Genes to cells : devoted to molecular & cellular mechanisms High 12059962
2004 AP-3-dependent trafficking controls ClC-3 sorting to synaptic vesicles. ClC-3 levels in synaptic vesicles and hippocampal mossy fiber terminals are reduced in AP-3-deficient mocha mice. In PC12 cells, ClC-3 traffics to synaptic-like microvesicles via the AP-3 route, and co-segregates with ZnT3, functionally increasing vesicular zinc transport. AP-3 deficient mouse model (mocha), subcellular fractionation, immunohistochemistry, brefeldin A trafficking assay, zinc transport assay The Journal of biological chemistry High 15073168
2004 CaMKII phosphorylates ClC-3 at serine 109 in the N-terminus in vitro, and an S109A mutation abolishes CaMKII-dependent Cl- conductance at the plasma membrane, establishing Ser109 as the critical CaMKII phosphorylation site for channel gating. In vitro CaMKII phosphorylation assay of ClC-3 N-terminus, S109A site-directed mutagenesis, whole-cell patch clamp in smooth muscle cells and HT29 cells The Journal of physiology High 14754994
2006 ClC-3 is required for NADPH oxidase (Nox1)-dependent ROS production and cytokine-induced NF-κB activation in vascular smooth muscle cells. ClC-3 co-localizes and co-immunoprecipitates with Nox1 and p22phox in early endosomes, and is required for charge neutralization of the electron flow generated by Nox1 across signaling endosome membranes. Co-immunoprecipitation, colocalization by confocal microscopy, ClC-3 knockdown, ROS measurement, NF-κB reporter assay in SMCs Circulation research High 17673675
2006 ClC-3 is required for NADPH oxidase activation during phagocytosis and the respiratory burst in neutrophils. ClC-3 protein localizes to secretory vesicles and secondary granules and redistributes to the phagosomal membrane upon stimulation. Clcn3-/- PMNs show markedly reduced NADPH oxidase activity and phagocytosis. Clcn3-/- mouse model, synchronized phagocytosis assay, immunolocalization, NADPH oxidase activity assay The Journal of biological chemistry High 16522634
2007 ClC-3 overexpression in HEK293T cells induces a Cl-/H+ antiport current with outward rectification. Mutation of the gating glutamate E224A eliminates proton coupling (reversal potential becomes fully Cl--dependent), identifying ClC-3 as a Cl-/H+ antiporter similar to ClC-4 and ClC-5. Whole-cell patch clamp, pH manipulation, E224A mutagenesis in HEK293T cells American journal of physiology. Cell physiology High 17977943
2007 ClC-3 traffics through the plasma membrane via interaction of an N-terminal dileucine acidic cluster (amino acids 13-19) with clathrin. Alanine substitution of this cluster abolished clathrin binding, reduced endocytosis, and increased surface expression. Pulse-chase showed ~25% of newly synthesized ClC-3 transiently inserts into the plasma membrane before rapid endocytosis (t1/2 ~9 min). Co-immunoprecipitation, GST pulldown, surface biotinylation, pulse-chase with [35S]methionine, mutagenesis, immunofluorescence microscopy The Journal of biological chemistry High 17652080
2007 ClC-3 overexpression in lysosomes increases organelle acidification and enhances resistance to etoposide by ~2-fold, demonstrating that ClC-3-mediated Cl- flux drives intracellular vesicle acidification relevant to drug sequestration. ClC-3 overexpression in BON and HEK293 cells, acridine orange staining for acidification, etoposide IC50 measurement, V-ATPase inhibition Molecular cancer therapeutics Medium 17363491
2008 ClC-3 localizes to the plasma membrane and mitotic spindle during M phase and is required for premitotic cytoplasmic condensation (PMC). ShRNA knockdown of ClC-3 reduces Cl- efflux, decreases PMC, and impairs DNA condensation, placing ClC-3-mediated Cl- efflux as a mechanistic driver of cell volume reduction during mitosis. Colocalization immunofluorescence, co-immunoprecipitation, patch-clamp at M phase, shRNA knockdown with time-lapse PMC assay The Journal of neuroscience High 18784301
2008 ClC-3 localizes to endosomes and synaptic-like microvesicles (not LDCVs) in adrenal chromaffin and pancreatic beta cells. Loss of ClC-3 decreases exocytosis of large dense-core vesicles, indicating an indirect role via an endosomal trafficking step rather than direct LDCV function. Immunohistochemistry with newly generated antibodies, subcellular fractionation, capacitance measurements, amperometry in Clcn3-/- mice The Journal of neuroscience High 18923035
2009 ClC-3 is required for insulin granule priming and insulin secretion. ClC-3 is expressed on insulin secretory granules, and Clcn3-/- beta cells show ~80% reduction in depolarization-evoked exocytosis and ~44% reduction in proton transport across granule membranes. Clcn3-/- mice, capacitance measurements of single beta cell exocytosis, proton transport assay, immunoblotting/immunostaining/negative immuno-EM on phogrin-EGFP-labeled LDCVs Cell metabolism High 19808023
2009 ClC-3 Cl-/H+ antiport becomes uncoupled at low extracellular pH (≤5.5), behaving as a pure anion channel at pH 4.0. The extracellular gating glutamate E224 and cysteine residues at the extracellular face are required for normal pH-dependent coupling. Patch clamp with pH manipulation, E224A mutagenesis, cysteine deletion mutant (C103_P130del), MTSES alkanethiolation, siRNA knockdown The Journal of biological chemistry High 19926787
2010 CaMKII directly interacts with and co-immunoprecipitates with ClC-3 in glioma cells and patient glioblastoma biopsies. CaMKII activation enhances ClC-3 Cl- currents 3-fold; this regulation is absent after ClC-3 shRNA knockdown. CaMKII-dependent ClC-3 activity facilitates glioma cell invasion. Co-immunoprecipitation, shRNA knockdown, whole-cell patch clamp with intracellular CaMKII infusion, invasion assay The Journal of biological chemistry High 20139089
2010 TNF-α activates swelling-activated Cl- current (ICl,swell) in vascular smooth muscle cells through a ClC-3-dependent mechanism requiring endosomal H2O2 production by Nox1. ClC-3 null VSMCs lack TNF-α-induced ICl,swell. Disruption of endosome trafficking (mutant Rab5 or Rab11) blocks the effect, placing ClC-3 endosomal ROS production upstream of ICl,swell activation. Perforated patch clamp in Clcn3-/- and WT VSMCs, catalase treatment, dominant-negative Rab5/Rab11 mutants, H2O2 application The Journal of biological chemistry High 20479003
2011 Presynaptic ClC-3 co-localizes with VGAT on inhibitory synaptic vesicles in hippocampal CA1 and is required for vesicle acidification and GABA loading. Clcn3-/- mice show decreased amplitude and frequency of miniature inhibitory postsynaptic currents, and Cl--induced acidification of inhibitory vesicles is markedly reduced. Hippocampal slice electrophysiology in Clcn3-/- mice, colocalization with VGAT, synaptic vesicle acidification assay Nature neuroscience High 21378974
2011 CaMKII-dependent activation of ClC-3 drives cytoplasmic Cl- efflux mediating premitotic condensation (PMC) during mitotic cell rounding in glioma cells. Knockdown of ClC-3 eliminates CaMKII-dependent Cl- currents in dividing cells and impedes PMC. Time-lapse microscopy, whole-cell patch clamp, shRNA knockdown, KN-93 CaMKII inhibition American journal of physiology. Cell physiology High 22049206
2012 ClC-3 deletion in hippocampal neurons increases glutamatergic synaptic vesicle volume, amplitude and frequency of mEPSCs, and release probability, suggesting ClC-3 normally limits glutamate loading and release probability of synaptic vesicles. Electron microscopy confirmed increased SV volumes in Clcn3-/- hippocampi. Whole-cell electrophysiology in Clcn3-/- hippocampal neurons, competitive AMPA receptor antagonist assay (γ-DGG), electron microscopy, paired pulse ratio Frontiers in cellular neuroscience High 24904288
2013 ClC-3 is an intracellular Cl-/H+ exchanger with large voltage-dependent nonlinear capacitance. An N-terminal retention signal keeps it intracellular; removal of this signal permits electrophysiological characterization. Mutation of the proton glutamate decreases transport but increases capacitance. ClC-3 is more specialized for capacitive non-transporting cycles than ClC-4 or ClC-5. Whole-cell electrophysiology, fluorescence pH measurements, proton glutamate mutagenesis in heterologous expression system with N-terminal mutation ACS chemical neuroscience High 23509947
2013 Bradykinin-induced chemotaxis in glioma cells requires CaMKII-mediated activation of ClC-3. Simultaneous Ca2+ imaging and patch clamp showed bradykinin elevates [Ca2+]i, activates CaMKII, and induces Cl- currents attributed to ClC-3. shRNA knockdown of ClC-3 inhibits Ca2+-activated Cl- currents and abolishes bradykinin-induced chemotaxis. Simultaneous fura-2 Ca2+ imaging and perforated patch clamp, CaMKII inhibitor KN-93, shRNA ClC-3 knockdown, Boyden chamber chemotaxis assay The Journal of neuroscience High 23345219
2014 ClC-3 interacts with voltage-dependent anion channel 1 (VDAC1) to regulate mitochondrial cytochrome c release and intestinal epithelial cell apoptosis. DSS treatment reduces the ClC-3-VDAC1 interaction. ClC-3-/- mice have increased susceptibility to colitis and impaired Paneth cell function. Co-immunoprecipitation of ClC-3 and VDAC1, Clcn3-/- mouse colitis model (DSS/TNBS), apoptosis pathway analysis Gut Medium 24440986
2015 Three ClC-3 splice variants (ClC-3a, ClC-3b, ClC-3c) differ in subcellular localization but not transport function. ClC-3a and ClC-3b localize to late endosomes/lysosomes via dileucine-like motifs; ClC-3c targets to recycling endosomes via a novel N-terminal isoleucine-proline (IP) motif. All isoforms mediate identical outwardly rectifying Cl-/H+ currents. Subcellular localization by confocal microscopy, N-terminal dileucine motif mutagenesis, whole-cell patch clamp of surface-expressed variants in HEK293T cells The Journal of biological chemistry High 26342074
2016 ROCK2 kinase phosphorylates ClC-3 at Thr532, and this phosphorylation is required for angiotensin II-induced Cl- current and VSMC migration. ClC-3 co-immunoprecipitates with ROCK2; T532D (phosphomimetic) potentiates and T532A abolishes the AngII-induced current and migration. Site-directed mutagenesis (T532D, T532A), co-immunoprecipitation, N/C-terminal truncation, ROCK2 siRNA, whole-cell patch clamp, VSMC migration assay British journal of pharmacology High 26562480
2017 ClC-3c splice variant targets ClC-4 to endosomal compartments by forming ClC-3-ClC-4 heterodimers. In Clcn3-/- astrocytes, ClC-4 is retained in the ER. High-resolution clear native gel electrophoresis showed ClC-3-ClC-4 heterodimers are more stable than ClC-4 homodimers, explaining ClC-3-dependent trafficking of ClC-4. Clear native gel electrophoresis, subcellular localization in Clcn3-/- astrocytes, co-expression experiments in HEK293T cells The Journal of biological chemistry High 28972156
2018 The external gating glutamate E224 and internal anion gate tyrosine Y572 interact to regulate ClC-3 Cl-/H+ coupling. Y572S removal dramatically increases transport current and impairs coupling; Y572F (loss of -OH) alters anion selectivity and impairs coupling. M531A mutation improves coupling efficiency. E224 and Y572 form a 'closed gate' maintaining coupling during the transport cycle. Multiple site-directed mutations (E224A, Y572S, Y572F, M531A), whole-cell patch clamp, cytoplasmic alkalization measurement, gating charge analysis The Journal of physiology High 29917234
2018 ClC-3 Cl-/H+ transporter regulates HER2 transcription in breast cancer cells through modulation of intracellular Cl- concentration, operating via the STAT3 signaling pathway. siRNA-mediated ClC-3 knockdown represses HER2 transcription and decreases STAT3 phosphorylation. siRNA knockdown of ClC-3, HER2 transcription assay, STAT3/AKT/mTOR phosphorylation Western blot Cancer science Low 29949674
2018 ClC-3 promotes Ang II-induced NADPH oxidase activation and ROS production in endothelial cells by facilitating Nox2/p22phox expression and p38 MAPK-dependent translocation of p47phox/p67phox to the membrane, increasing Nox2 complex assembly. siRNA knockdown and overexpression of ClC-3, NADPH oxidase activity assay, p47phox/p67phox membrane translocation assay, p38 MAPK inhibitor, ROS measurement Acta pharmacologica Sinica Medium 29977005
2007 The short ClC-3 isoform (sClC-3) C-terminal cytoplasmic tail directly binds filamentous (F-)actin but not globular actin in co-sedimentation assays. The F-actin binding region maps to amino acids 690-760. Dialysis of synthetic peptides blocking this interaction into cells reduced swelling-activated Cl- current by 38-60%, linking direct actin interaction to VSOAC channel activity. GST co-sedimentation assay with F-actin and G-actin, truncation mapping, synthetic peptide dialysis in NIH/3T3 cells, patch clamp The Journal of biological chemistry High 17442672
2008 Ins(3,4,5,6)P4 inhibits ClC-3 Cl- conductance both at the plasma membrane in HEK cells and in early endosomes (measured by intra-endosomal pH via FITC-transferrin), and inhibits endogenous ClC-3 conductance in postsynaptic membranes of hippocampal neurons, establishing Ins(3,4,5,6)P4 as an endogenous ClC-3 regulator. Whole-cell patch clamp of heterologous ClC-3, fluorescence ratio imaging of endosomal pH via FITC-transferrin, electrophysiology in hippocampal neurons Current biology : CB High 18951024
2002 ClC-3B splice variant contains a PDZ-binding motif and interacts with EBP50 scaffold protein in vitro and in vivo. Co-transfection of ClC-3B with EBP50 recruits ClC-3B to plasma membrane ruffles and induces outwardly rectifying Cl- channel (ORCC) activity at membrane ruffles that can be activated via PKA when CFTR is also co-expressed. In vitro and in vivo binding assays, patch clamp at membrane ruffles, C127 cell transfection with ClC-3B and EBP50 FASEB journal Medium 11967229
2016 Endophilin A2 interacts with ClC-3 via its SH3 domain and promotes ClC-3 transport from the cytoplasm to the cell membrane in VSMCs, increasing volume-regulated Cl- current. Co-immunoprecipitation and live cell imaging confirmed this trafficking interaction. Co-immunoprecipitation, live cell imaging, Western blot in endophilin A2 transgenic mice, whole-cell patch clamp Circulation journal Medium 27760895
2012 CLC-3 channels at postsynaptic sites associate with NMDA receptors and moderate long-term potentiation (LTP) at Schaffer collateral-CA1 synapses. Loss of CLC-3 increases LTP by ~40%. A decoy peptide for the CaMKII phosphorylation site on CLC-3 blocks its regulatory function in LTP, demonstrating phosphorylation is required. Hippocampal slice LTP recordings in Clcn3-/- mice, CaMKII phosphorylation site decoy peptide, coimmunoprecipitation of ClC-3 with NMDA receptors The Journal of physiology High 23165767
2006 CLC-3 is expressed on the plasma membrane at postsynaptic sites and co-immunoprecipitates with NMDA receptors in neonatal hippocampal neurons. CaMKII-activated CLC-3 Cl- conductance is absent in clc-3-/- mice and enhances the decay time of NMDA receptor-mediated mEPSPs in a Cl--dependent manner, demonstrating that CLC-3 modulates excitatory synaptic strength. Surface biotinylation, immunohistochemistry, electron microscopy, co-immunoprecipitation, whole-cell patch clamp in clc-3-/- mice, Cl- dialysis Neuron High 17046694
2021 De novo heterozygous CLCN3 missense variants (e.g., p.Ile607Thr and p.Thr570Ile) cause gain-of-function changes: increased Cl-/H+ exchange currents at negative voltages and loss of inhibition by luminal acidic pH, leading to neurodevelopmental disorders. Homozygous loss-of-function variants cause severe neurodegeneration resembling the Clcn3-/- mouse phenotype. Electrophysiology in Xenopus oocytes and mammalian cells expressing patient variants, patient MRI/clinical phenotyping American journal of human genetics High 34186028
2001 Tissue-specific N-glycosylation of ClC-3 accounts for differential electrophoretic mobility across brain, intestine, and kidney; all isoforms resolve to the same molecular mass after enzymatic removal of N-linked oligosaccharides. Western blot after enzymatic N-glycosidase digestion of membrane proteins from multiple mouse tissues Biochemical and biophysical research communications Medium 11511107
2012 ClC-3 subcellular localization changes in a cell cycle-dependent manner in HeLa cells: nuclear in early/late G1 and S phase, cytoplasmic in G2, then redistributes to spindle poles and between chromosomes during mitosis, indicating nucleocytoplasmic shuttling linked to cell cycle stage. Immunofluorescence confocal microscopy with cell cycle synchronization in HeLa cells Histochemistry and cell biology Medium 22371056

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2007 Cytokine activation of nuclear factor kappa B in vascular smooth muscle cells requires signaling endosomes containing Nox1 and ClC-3. Circulation research 174 17673675
1999 A serine residue in ClC-3 links phosphorylation-dephosphorylation to chloride channel regulation by cell volume. The Journal of general physiology 148 9874688
2001 Regulation of human CLC-3 channels by multifunctional Ca2+/calmodulin-dependent protein kinase. The Journal of biological chemistry 127 11274166
2006 Anion channels, including ClC-3, are required for normal neutrophil oxidative function, phagocytosis, and transendothelial migration. The Journal of biological chemistry 124 16522634
2002 CLC-3 deficiency leads to phenotypes similar to human neuronal ceroid lipofuscinosis. Genes to cells : devoted to molecular & cellular mechanisms 123 12059962
2002 The ClC-3 chloride channel promotes acidification of lysosomes in CHO-K1 and Huh-7 cells. American journal of physiology. Cell physiology 115 11997263
2010 Molecular interaction and functional regulation of ClC-3 by Ca2+/calmodulin-dependent protein kinase II (CaMKII) in human malignant glioma. The Journal of biological chemistry 110 20139089
2000 Biophysical properties of ClC-3 differentiate it from swelling-activated chloride channels in Chinese hamster ovary-K1 cells. The Journal of biological chemistry 109 10973952
2004 AP-3-dependent mechanisms control the targeting of a chloride channel (ClC-3) in neuronal and non-neuronal cells. The Journal of biological chemistry 98 15073168
2000 The role of ClC-3 in volume-activated chloride currents and volume regulation in bovine epithelial cells demonstrated by antisense inhibition. The Journal of physiology 98 10747184
2008 ClC3 is a critical regulator of the cell cycle in normal and malignant glial cells. The Journal of neuroscience : the official journal of the Society for Neuroscience 97 18784301
2002 ClC-3 is a fundamental molecular component of volume-sensitive outwardly rectifying Cl- channels and volume regulation in HeLa cells and Xenopus laevis oocytes. The Journal of biological chemistry 95 12183454
2002 Deficiency in ClC-3 chloride channels prevents rat aortic smooth muscle cell proliferation. Circulation research 94 12433844
2006 The ClC-3 Cl- channel in cell volume regulation, proliferation and apoptosis in vascular smooth muscle cells. Trends in pharmacological sciences 90 16697056
2002 Altered GABAergic function accompanies hippocampal degeneration in mice lacking ClC-3 voltage-gated chloride channels. Brain research 90 12470859
2001 Human ClC-3 is not the swelling-activated chloride channel involved in cell volume regulation. The Journal of biological chemistry 85 11278960
2004 Regulation of intracellular Cl- concentration through volume-regulated ClC-3 chloride channels in A10 vascular smooth muscle cells. The Journal of biological chemistry 84 15596438
2002 Secretion and cell volume regulation by salivary acinar cells from mice lacking expression of the Clcn3 Cl- channel gene. The Journal of physiology 83 12433961
2013 Bradykinin-induced chemotaxis of human gliomas requires the activation of KCa3.1 and ClC-3. The Journal of neuroscience : the official journal of the Society for Neuroscience 81 23345219
2013 ClC-3 is an intracellular chloride/proton exchanger with large voltage-dependent nonlinear capacitance. ACS chemical neuroscience 75 23509947
2010 Invasion of human glioma cells is regulated by multiple chloride channels including ClC-3. Anticancer research 75 21115901
2007 Overexpression of CLC-3 in HEK293T cells yields novel currents that are pH dependent. American journal of physiology. Cell physiology 75 17977943
1995 Characterization of a human and murine gene (CLCN3) sharing similarities to voltage-gated chloride channels and to a yeast integral membrane protein. Genomics 71 7665160
2008 Suppression of ClC-3 channel expression reduces migration of nasopharyngeal carcinoma cells. Biochemical pharmacology 69 18359479
2003 Functional effects of novel anti-ClC-3 antibodies on native volume-sensitive osmolyte and anion channels in cardiac and smooth muscle cells. American journal of physiology. Heart and circulatory physiology 69 12816749
2002 ClC-3B, a novel ClC-3 splicing variant that interacts with EBP50 and facilitates expression of CFTR-regulated ORCC. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 69 11967229
2001 Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody. The Journal of physiology 69 11230516
2000 Expression and canalicular localization of two isoforms of the ClC-3 chloride channel from rat hepatocytes. American journal of physiology. Gastrointestinal and liver physiology 67 10915634
2008 Silence of ClC-3 chloride channel inhibits cell proliferation and the cell cycle via G/S phase arrest in rat basilar arterial smooth muscle cells. Cell proliferation 65 18823498
2006 CLC-3 channels modulate excitatory synaptic transmission in hippocampal neurons. Neuron 65 17046694
2005 ClC-3 chloride channel is upregulated by hypertrophy and inflammation in rat and canine pulmonary artery. British journal of pharmacology 65 15723096
2004 Bcl-2-dependent modulation of swelling-activated Cl- current and ClC-3 expression in human prostate cancer epithelial cells. Cancer research 65 15256454
1998 The swelling-activated chloride channel ClC-2, the chloride channel ClC-3, and ClC-5, a chloride channel mutated in kidney stone disease, are expressed in distinct subpopulations of renal epithelial cells. The Journal of clinical investigation 65 9449697
1996 Association of ClC-3 channel with Cl- transport by human nonpigmented ciliary epithelial cells. The Journal of membrane biology 65 8661780
2011 The ClC-3 chloride channels in cardiovascular disease. Acta pharmacologica Sinica 60 21602838
2004 ClC-3-independent, PKC-dependent activity of volume-sensitive Cl channel in mouse ventricular cardiomyocytes. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 60 15319524
2010 Activation of swelling-activated chloride current by tumor necrosis factor-alpha requires ClC-3-dependent endosomal reactive oxygen production. The Journal of biological chemistry 59 20479003
2004 Identification of an N-terminal amino acid of the CLC-3 chloride channel critical in phosphorylation-dependent activation of a CaMKII-activated chloride current. The Journal of physiology 58 14754994
2008 Role of the vesicular chloride transporter ClC-3 in neuroendocrine tissue. The Journal of neuroscience : the official journal of the Society for Neuroscience 56 18923035
2011 Presynaptic CLC-3 determines quantal size of inhibitory transmission in the hippocampus. Nature neuroscience 55 21378974
2003 Fundamental role of ClC-3 in volume-sensitive Cl- channel function and cell volume regulation in AGS cells. American journal of physiology. Gastrointestinal and liver physiology 53 12842831
2015 Neuronal ClC-3 Splice Variants Differ in Subcellular Localizations, but Mediate Identical Transport Functions. The Journal of biological chemistry 52 26342074
2013 Discovery of bufadienolides as a novel class of ClC-3 chloride channel activators with antitumor activities. Journal of medicinal chemistry 51 23799775
2009 The ClC-3 Cl-/H+ antiporter becomes uncoupled at low extracellular pH. The Journal of biological chemistry 51 19926787
2007 ClC-3 is required for LPA-activated Cl- current activity and fibroblast-to-myofibroblast differentiation. American journal of physiology. Cell physiology 51 18077605
2014 CLC-3 channels in cancer (review). Oncology reports 50 25421907
2000 Molecular distribution of volume-regulated chloride channels (ClC-2 and ClC-3) in cardiac tissues. American journal of physiology. Heart and circulatory physiology 49 11045957
2007 ClC-3 expression enhances etoposide resistance by increasing acidification of the late endocytic compartment. Molecular cancer therapeutics 47 17363491
2014 ClC-3 chloride channel/antiporter defect contributes to inflammatory bowel disease in humans and mice. Gut 46 24440986
2009 Suppression of sulfonylurea- and glucose-induced insulin secretion in vitro and in vivo in mice lacking the chloride transport protein ClC-3. Cell metabolism 45 19808023
2008 ClC-3 and IClswell are required for normal neutrophil chemotaxis and shape change. The Journal of biological chemistry 44 18840613
2007 The ClC-3 chloride transport protein traffics through the plasma membrane via interaction of an N-terminal dileucine cluster with clathrin. The Journal of biological chemistry 44 17652080
2007 Endotoxin priming of neutrophils requires NADPH oxidase-generated oxidants and is regulated by the anion transporter ClC-3. The Journal of biological chemistry 42 17908687
2006 ClC-3 chloride channel prevents apoptosis induced by thapsigargin in PC12 cells. Apoptosis : an international journal on programmed cell death 41 16520896
2014 The impact of a chlorotoxin-modified liposome system on receptor MMP-2 and the receptor-associated protein ClC-3. Biomaterials 39 24743031
2021 Unique variants in CLCN3, encoding an endosomal anion/proton exchanger, underlie a spectrum of neurodevelopmental disorders. American journal of human genetics 37 34186028
2011 ClC-3 chloride channel prevents apoptosis induced by hydrogen peroxide in basilar artery smooth muscle cells through mitochondria dependent pathway. Apoptosis : an international journal on programmed cell death 37 21373935
2010 ClC-3 chloride channels are essential for cell proliferation and cell cycle progression in nasopharyngeal carcinoma cells. Acta biochimica et biophysica Sinica 37 20539936
2008 Intracellular ClC-3 chloride channels promote bone resorption in vitro through organelle acidification in mouse osteoclasts. American journal of physiology. Cell physiology 37 18234851
2016 ClC-3 Chloride Channel Proteins Regulate the Cell Cycle by Up-regulating cyclin D1-CDK4/6 through Suppressing p21/p27 Expression in Nasopharyngeal Carcinoma Cells. Scientific reports 35 27451945
2009 Cardiac-specific, inducible ClC-3 gene deletion eliminates native volume-sensitive chloride channels and produces myocardial hypertrophy in adult mice. Journal of molecular and cellular cardiology 35 19615374
2013 Acid-sensitive outwardly rectifying (ASOR) anion channels in human epithelial cells are highly sensitive to temperature and independent of ClC-3. Pflugers Archiv : European journal of physiology 34 23708799
2014 Involvement of ClC-3 chloride/proton exchangers in controlling glutamatergic synaptic strength in cultured hippocampal neurons. Frontiers in cellular neuroscience 33 24904288
2016 Abrogating ClC-3 Inhibits LPS-induced Inflammation via Blocking the TLR4/NF-κB Pathway. Scientific reports 31 27363391
2013 The ClC-3 chloride channel associated with microtubules is a target of paclitaxel in its induced-apoptosis. Scientific reports 31 24026363
2005 ClC-3-independent sensitivity of apoptosis to Cl- channel blockers in mouse cardiomyocytes. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 31 16037691
2004 Hyposmotic activation of ICl,swell in rabbit nonpigmented ciliary epithelial cells involves increased ClC-3 trafficking to the plasma membrane. Biochemistry and cell biology = Biochimie et biologie cellulaire 31 15674438
2013 ClC-3 deficiency prevents apoptosis induced by angiotensin II in endothelial progenitor cells via inhibition of NADPH oxidase. Apoptosis : an international journal on programmed cell death 30 23873092
2012 ClC-3 is a candidate of the channel proteins mediating acid-activated chloride currents in nasopharyngeal carcinoma cells. American journal of physiology. Cell physiology 29 22496242
2012 Cell cycle-dependent subcellular distribution of ClC-3 in HeLa cells. Histochemistry and cell biology 28 22371056
2011 Kinase activation of ClC-3 accelerates cytoplasmic condensation during mitotic cell rounding. American journal of physiology. Cell physiology 28 22049206
2001 Tissue-specific N-glycosylation of the ClC-3 chloride channel. Biochemical and biophysical research communications 28 11511107
2011 Ion-deficient environment induces the expression of basolateral chloride channel, ClC-3-like protein, in gill mitochondrion-rich cells for chloride uptake of the tilapia Oreochromis mossambicus. Physiological and biochemical zoology : PBZ 27 21091354
2010 Chloride channel ClC-3 in gills of the euryhaline teleost, Tetraodon nigroviridis: expression, localization and the possible role of chloride absorption. The Journal of experimental biology 27 20154183
2010 Chloride channel ClC-3 promotion of osteogenic differentiation through Runx2. Journal of cellular biochemistry 27 20506205
2008 An expanded biological repertoire for Ins(3,4,5,6)P4 through its modulation of ClC-3 function. Current biology : CB 27 18951024
2017 Suppression of CLC-3 chloride channel reduces the aggressiveness of glioma through inhibiting nuclear factor-κB pathway. Oncotarget 26 28969029
2005 Inhibition of swelling-activated Cl- currents by functional anti-ClC-3 antibody in native bovine non-pigmented ciliary epithelial cells. Investigative ophthalmology & visual science 25 15728552
2018 Modulation of ClC-3 gating and proton/anion exchange by internal and external protons and the anion selectivity filter. The Journal of physiology 24 29917234
2017 Activation of ClC-3 chloride channel by 17β-estradiol relies on the estrogen receptor α expression in breast cancer. Journal of cellular physiology 24 28419445
2017 Preferential association with ClC-3 permits sorting of ClC-4 into endosomal compartments. The Journal of biological chemistry 24 28972156
2022 HNRNPK/CLCN3 axis facilitates the progression of LUAD through CAF-tumor interaction. International journal of biological sciences 23 36439880
2019 Antitumor effects of disulfiram/copper complex in the poorly-differentiated nasopharyngeal carcinoma cells via activating ClC-3 chloride channel. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 23 31606620
2018 Transcriptional repression of human epidermal growth factor receptor 2 by ClC-3 Cl- /H+ transporter inhibition in human breast cancer cells. Cancer science 23 29949674
2014 Swelling-activated Cl- currents and intracellular CLC-3 are involved in proliferation of human pulmonary artery smooth muscle cells. Journal of hypertension 23 24284495
2011 ClC-3 is a main component of background chloride channels activated under isotonic conditions by autocrine ATP in nasopharyngeal carcinoma cells. Journal of cellular physiology 23 21792908
1999 Single-cell RT-PCR demonstrates expression of voltage-dependent chloride channels (ClC-1, ClC-2 and ClC-3) in outer hair cells of rat cochlea. Brain research 23 10446329
2006 Single-channel properties of volume-sensitive Cl- channel in ClC-3-deficient cardiomyocytes. The Japanese journal of physiology 22 16441975
2004 ClC-3 expression in the cell cycle of nasopharyngeal carcinoma cells. Sheng li xue bao : [Acta physiologica Sinica] 22 15127135
2018 ClC-3 promotes angiotensin II-induced reactive oxygen species production in endothelial cells by facilitating Nox2 NADPH oxidase complex formation. Acta pharmacologica Sinica 21 29977005
2015 ClC-3 chloride channel modulates the proliferation and migration of osteosarcoma cells via AKT/GSK3β signaling pathway. International journal of clinical and experimental pathology 21 25973047
2012 Functional regulation of ClC-3 in the migration of vascular smooth muscle cells. Hypertension (Dallas, Tex. : 1979) 21 23150504
2016 Threonine532 phosphorylation in ClC-3 channels is required for angiotensin II-induced Cl(-) current and migration in cultured vascular smooth muscle cells. British journal of pharmacology 20 26562480
2013 Deficiency of volume-regulated ClC-3 chloride channel attenuates cerebrovascular remodelling in DOCA-salt hypertension. Cardiovascular research 20 23786998
2007 Hypotonic activation of short ClC3 isoform is modulated by direct interaction between its cytosolic C-terminal tail and subcortical actin filaments. The Journal of biological chemistry 20 17442672
2016 ClC-3 Expression and Its Association with Hyperglycemia Induced HT22 Hippocampal Neuronal Cell Apoptosis. Journal of diabetes research 19 26925421
2016 Endophilin A2 Influences Volume-Regulated Chloride Current by Mediating ClC-3 Trafficking in Vascular Smooth Muscle Cells. Circulation journal : official journal of the Japanese Circulation Society 19 27760895
2014 A newly cloned ClC-3 isoform, ClC-3d, as well as ClC-3a mediates Cd-sensitive outwardly rectifying anion currents. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 19 24603049
2014 ClC-3 deficiency protects preadipocytes against apoptosis induced by palmitate in vitro and in type 2 diabetes mice. Apoptosis : an international journal on programmed cell death 19 25218423
2012 CLC-3 chloride channels moderate long-term potentiation at Schaffer collateral-CA1 synapses. The Journal of physiology 19 23165767

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