| Annotated protein: | H(+)/Cl(-) exchange transporter 3 (Chloride channel protein 3) (ClC-3) (Chloride transporter ClC-3). Gene symbol: CLCN3. Taxonomy: Homo sapiens (Human). Uniprot ID: P51790-1 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ CLCN3 |
| Ontology domain: | Cellular Component |
| SynGO term: | integral component of synaptic vesicle membrane (GO:0030285) |
| Synapse type(s): | hippocampus brain, glutamatergic brain, GABAergic |
| Annotated paper: | Stobrawa SM, et al. "Disruption of ClC-3, a chloride channel expressed on synaptic vesicles, leads to a loss of the hippocampus" Neuron. 2001 Jan;29(1):185-96 PMID:11182090 |
| Figure(s): | Fig. 7 E, D |
| Annotation description: | A human HA-tagged ClC-3 was overexpressed in rat hippocampal neurons and its colocalization with synaptic vesicle marker protein synaptophysin was detected by immunofluorescence (Fig. 7E). Additionally, by means of biochemical fractionation and Western blotting, the authors of the study showed that ClC-3 is expressed on glutamatergic and GABAergic synaptic vesicles (isolated from rat brain) (Fig. 7 B-C). The data suggests indirectly that ClC3 localizes to synaptic vesicles. A FLAG-tagged human ClC-3 was expressed in COS-7 cells and chloride channel co-localization with lamp-1 positive puncta was assessed by immunofluorescence (Fig. 7 D). This data supports the endosomal localization of ClC-3 in these cells, however no direct evidence for the localization of human ClC-3 to presynaptic endosomes is shown. 6/2/2018 Pim - Based on the known transmembrane topology of the protein, the "integral component of ... membrane" term was chosen. |
| Evidence tracking, Biological System: | Intact tissue Cultured neurons |
| Evidence tracking, Protein Targeting: | Over-expression Antibody (detection) |
| Evidence tracking, Experiment Assay: | Confocal Western blot Biochemical fractionation (generic) |
| Annotator(s): | Momchil Ninov (ORCID:0000-0002-0808-7003) Mahdokht Kohansalnodehi (ORCID:0000-0002-3898-5197) Reinhard Jahn (ORCID:0000-0003-1542-3498) |
| Lab: | Department of Neurobiology, Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany |
| Additional literature: | Suppl Fig 1-2 suggest indirectly localization of ClC-3 to synaptic vesicles (combination of subcellular fractionation of rat brain and immunoblotting) @ PMID:19169251 Guzman and et al (Fig 6 A-C; Fig 3B) have identified in mouse brain different isoforms of ClC-3. These isoforms' cDNAs (cDNAs encoding full-length mouse ClC-3a, ClC-3b, or ClC-3c (GenBankTM Accession Number NM_007711.3, NM_173873.1, NM_173876.3) were fused in-frame to the 5 -end of the coding sequences of enhanced green or monomeric red fluorescence protein (eGFP or mRFP) and cloned into FsY1.1 G.W. or p156rrL vectors. The authors show by heterologous expression of mouse brain ClC-3e isoform and co-expression with either transferrin receptor (TfR) or Rab11 that the protein isoform ClC-3c is localized both on plasma membrane and TfR- and Rab11-positive endosomes. Furthermore, Guzman et al show co-localization of mouse brain ClC-3a isoform with Lamp-1 (Fig. 3B) and Rab 7 (Fig 6A) by immunofluorescence and suggested late endosomal/lysosomal expression of the protein (same findings also for heterologous expression of the protein in MDCK cells, data not shown in the paper), No direct evidence is provided for similar subcellular localization of the protein isoform ClC-3a in neurons. eGFP-tagged mouse brain ClC-3b (ClC-3A) protein isoform was heterologously expressed in HEK293T cells. Subcellular localization was examined by immunostaining for organellar markers LAMP1 (Fig. 3B) and Rab 11 and rab 7 (Fig. 6B). The data suggested late endosomal /lysosomal expression of the chloride channel. The data is supported by co-immunostaining of human ClC-3b with lamp-1 in COS-7 cells and copurification of mouse ClC-3b with lamp-1 and rab4 (Stobrawa, S. et al, Neuron, 2001). However, no direct evidence for this protein isoform expression at synaptic endosomes is shown. Overall, despite the initial attempt to characterize these isoforms by exogeneous expressing in cell lines, further investigation of their molecular functions in neurons is required. @ PMID:26342074 |
| SynGO annotation ID: | 366 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |