Ion permeation through a Cl--selective channel designed from a CLC Cl-/H+ exchanger

Proc Natl Acad Sci U S A. 2008 Aug 12;105(32):11194-9. doi: 10.1073/pnas.0804503105. Epub 2008 Aug 4.

Abstract

The CLC family of Cl(-)-transporting proteins includes both Cl(-) channels and Cl(-)/H(+) exchange transporters. CLC-ec1, a structurally known bacterial homolog of the transporter subclass, exchanges two Cl(-) ions per proton with strict, obligatory stoichiometry. Point mutations at two residues, Glu(148) and Tyr(445), are known to impair H(+) movement while preserving Cl(-) transport. In the x-ray crystal structure of CLC-ec1, these residues form putative "gates" flanking an ion-binding region. In mutants with both of the gate-forming side chains reduced in size, H(+) transport is abolished, and unitary Cl(-) transport rates are greatly increased, well above values expected for transporter mechanisms. Cl(-) transport rates increase as side-chain volume at these positions is decreased. The crystal structure of a doubly ungated mutant shows a narrow conduit traversing the entire protein transmembrane width. These characteristics suggest that Cl(-) flux through uncoupled, ungated CLC-ec1 occurs via a channel-like electrodiffusion mechanism rather than an alternating-exposure conformational cycle that has been rendered proton-independent by the gate mutations.

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Chloride Channels / chemistry*
  • Chloride Channels / genetics
  • Chloride Channels / metabolism
  • Chlorides / chemistry*
  • Chlorides / metabolism
  • Crystallography, X-Ray
  • Humans
  • Ion Transport / physiology
  • Point Mutation
  • Protein Structure, Tertiary / genetics
  • Protons*

Substances

  • Chloride Channels
  • Chlorides
  • Protons

Associated data

  • PDB/3DET