Novel insights into the structural basis of pH-sensitivity in inward rectifier K+ channels Kir2.3

Cell Physiol Biochem. 2008;21(5-6):347-56. doi: 10.1159/000129629. Epub 2008 Apr 24.

Abstract

The Kir2 channels belong to a family of potassium selective channels with characteristic strong inward rectification. Heteromeric assemblies of Kir2.1, Kir2.2 and Kir2.3 channels underly membrane potential stabilizing currents in ventricular myocytes, neurons and skeletal muscle. Kir2 channels differ substantially in their sensitivity to extracellular pH. The extracellular histidine Kir2.3(H117) contributes to the pH dependence of K-channels containing Kir2.3. Here, we study the possibility of intramolecular interactions of the residue Kir2.3(H117) with conserved cysteines in close proximity to the selectivity filter. We engineered a cobalt coordination site and reduction/oxidation sensitivity in Kir2.3 by introduction of a cysteine into the putatively hydrogen bonding residue (Kir2.3(H117C)) confirming that this residue is in proximity to Kir2.3(C141). Using SCAM we determined the location of the Kir2.3(H117) in the outer pore mouth and incorporated these data into a 3D model. We conclude that formation of a hydrogen bond at low pH may stabilize the outer pore domain to favour the selectivity filter in a slightly distorted conformation thus reducing ion permeation. The data provide molecular insight into the unique pH regulation of inward rectifier channels.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cobalt / pharmacology
  • Disulfides / metabolism
  • Electrophysiology
  • Female
  • Humans
  • Hydrogen-Ion Concentration
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation / genetics
  • Oocytes / drug effects
  • Oocytes / metabolism
  • Patch-Clamp Techniques
  • Potassium Channels, Inwardly Rectifying / chemistry*
  • Potassium Channels, Inwardly Rectifying / genetics
  • Potassium Channels, Inwardly Rectifying / metabolism*
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Structural Homology, Protein
  • Substrate Specificity
  • Xenopus laevis

Substances

  • Disulfides
  • KCNJ4 protein, human
  • Potassium Channels, Inwardly Rectifying
  • Cobalt