Functional characterization of recombinant human CIC-4 chloride channels in cultured mammalian cells

Carlos G. Vanoye, Alfred L. George*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

48 Scopus citations


Members of the ClC chloride channel family participate in several physiological processes and are linked to human genetic diseases. The physiological role of ClC-4 is unknown and previous detailed characterizations of recombinant human ClC-4 (hClC-4) have provided conflicting results. To reexamine the hClC-4 phenotype, recombinant hClC-4 was expressed in three distinct mammalian cell lines and characterized using patch-clamp techniques. In all cells, the expression of hClC-4 generated strongly outward-rectifying Cl- currents with the conductance sequence: SCN- ≫ NO3- ≫ Cl- > Br- ≈ I- ≫ aspartate. Continuous activity of hClC-4 was sustained to different degrees by internal nucleotides: ATP ≈ ATPγS ≫ AMP-PNP ≈ GTP > ADP. Although non-hydrolysable nucleotides are sufficient for channel function, ATP hydrolysis is required for full activity. Changing the extracellular (2 mM or nominal Caa+-free) or intracellular Ca2+ (25 or 250 nM) concentration did not alter hClC-4 currents. Acidification of external pH (pHo) inhibited hClC-4 currents (half-maximal inhibition ≈ 6.19), whereas neither external alkalinization to pH 8.4 nor internal acidification to pH 6.0 reduced current levels. Single-channel recordings demonstrated a Cl- channel active only at depolarizing potentials with a slope conductance of ∼3 pS. Acidic pHo did not alter single-channel conductance. We conclude that recombinant hClC-4 encodes a small-conductance, nucleotide-dependent, Ca2+ -independent outward-rectifying chloride channel that is inhibited by external acidification. This detailed characterization will be highly valuable in comparisons of hClC-4 function with native chloride channel activities and for future structure-function correlations.

Original languageEnglish (US)
Pages (from-to)373-383
Number of pages11
JournalJournal of physiology
Issue number2
StatePublished - Mar 1 2002

ASJC Scopus subject areas

  • Physiology


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