Toxin-induced conformational changes in a potassium channel revealed by solid-state NMR

Nature. 2006 Apr 13;440(7086):959-62. doi: 10.1038/nature04649.

Abstract

The active site of potassium (K+) channels catalyses the transport of K+ ions across the plasma membrane--similar to the catalytic function of the active site of an enzyme--and is inhibited by toxins from scorpion venom. On the basis of the conserved structures of K+ pore regions and scorpion toxins, detailed structures for the K+ channel-scorpion toxin binding interface have been proposed. In these models and in previous solution-state nuclear magnetic resonance (NMR) studies using detergent-solubilized membrane proteins, scorpion toxins were docked to the extracellular entrance of the K+ channel pore assuming rigid, preformed binding sites. Using high-resolution solid-state NMR spectroscopy, here we show that high-affinity binding of the scorpion toxin kaliotoxin to a chimaeric K+ channel (KcsA-Kv1.3) is associated with significant structural rearrangements in both molecules. Our approach involves a combined analysis of chemical shifts and proton-proton distances and demonstrates that solid-state NMR is a sensitive method for analysing the structure of a membrane protein-inhibitor complex. We propose that structural flexibility of the K+ channel and the toxin represents an important determinant for the high specificity of toxin-K+ channel interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / drug effects
  • Bacterial Proteins / metabolism
  • Kv1.3 Potassium Channel / chemistry
  • Kv1.3 Potassium Channel / drug effects
  • Kv1.3 Potassium Channel / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Nuclear Magnetic Resonance, Biomolecular*
  • Oocytes / metabolism
  • Potassium Channel Blockers / chemistry
  • Potassium Channel Blockers / metabolism
  • Potassium Channel Blockers / pharmacology
  • Potassium Channels / chemistry*
  • Potassium Channels / drug effects*
  • Potassium Channels / metabolism
  • Protein Conformation / drug effects
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / drug effects
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Scorpion Venoms / chemistry
  • Scorpion Venoms / genetics
  • Scorpion Venoms / metabolism
  • Scorpion Venoms / pharmacology*
  • Scorpions
  • Xenopus

Substances

  • Bacterial Proteins
  • Kv1.3 Potassium Channel
  • Potassium Channel Blockers
  • Potassium Channels
  • Recombinant Fusion Proteins
  • Scorpion Venoms
  • kaliotoxin
  • prokaryotic potassium channel