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New Insights into the Mechanism of NO₃ ^- Selectivity in the Human Kidney Chloride Channel ClC-Ka and the CLC Protein Family.

ABSTRACT:

Background

The mechanism of anion selectivity in the human kidney chloride channels ClC-Ka and ClC-Kb is unknown. However, it has been thought to be very similar to that of other channels and antiporters of the CLC protein family, and to rely on anions interacting with a conserved Ser residue (Ser_cen) at the center of three anion binding sites in the permeation pathway S_cen. In both CLC channels and antiporters, mutations of Ser_cen alter the anion selectivity. Structurally, the side chain of Ser_cen of CLC channels and antiporters typically projects into the pore and coordinates the anion bound at S_cen.

Methods

To investigate the role of several residues in anion selectivity of ClC-Ka, we created mutations that resulted in amino acid substitutions in these residues. We also used electrophysiologic techniques to assess the properties of the mutants.

Results

Mutations in ClC-Ka that change Ser_cen to Gly, Pro, or Thr have only minor effects on anion selectivity, whereas the mutations in residues Y425A, F519A, and Y520A increase the NO₃ ^-/Cl^- permeability ratio, with Y425A having a particularly strong effect.

Conclusion

s ClC-Ka's mechanism of anion selectivity is largely independent of Ser_cen, and it is therefore unique in the CLC protein family. We identified the residue Y425 in ClC-Ka-and the corresponding residue (A417) in the chloride channel ClC-0-as residues that contribute to NO₃ ^- discrimination in these channels. This work provides important and timely insight into the relationship between structure and function for the kidney chloride channels ClC-Ka and ClC-Kb, and for CLC proteins in general.

SUBMITTER: Lagostena L

PROVIDER: S-EPMC6362620 | biostudies-literature |

REPOSITORIES: biostudies-literature

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