Modelling of the III-IV loop, a domain involved in calcium channel Ca(v)2.1 inactivation, highlights a structural homology with the gamma subunit of G proteins

Eur J Neurosci. 2002 Jul;16(2):219-28. doi: 10.1046/j.1460-9568.2002.02074.x.

Abstract

We have modelled the conformation of the III-IV loop of the Ca(v)2.1 subunit of P/Q calcium channels, a loop that is implicated in fast voltage-dependent inactivation. Change in channel inactivation requires its direct interaction with the I-II loop. This interaction occurs with an affinity in the order of 70 nm. Intracellular injection of a 40-mer III-IV loop-derived peptide produces an increase in the rate of fast inactivation. This alteration in channel kinetic is also accompanied by a hyperpolarizing shift in the steady-state voltage-dependence of inactivation. None of these effects are observed in the presence of a beta subunit, suggesting the existence of a competitive mechanism of action between the beta subunit and the III-IV loop. Amino acid sequence comparison using BLAST reveals that the III-IV loop shares 53% identity with the gamma subunit of G proteins. Because of the pivotal contribution of the III-IV loop to inactivation, an atomic model of the III-IV loop was generated by both homology modelling and molecular mechanics calculations. Using the X-ray structures of the betagamma dimer of the heterotrimeric G-proteins as templates, the III-IV loop is predicted to contain a well-structured alpha-helix at the amino-terminus with both the N- and C-termini having the same orientation in the plane of the inner lipid bilayer. We provide a hypothetical working model in which we propose that the III-IV loop interacts with the I-II loop via its Gbetagamma binding domain.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence / drug effects
  • Amino Acid Sequence / physiology
  • Animals
  • Binding Sites / physiology
  • Calcium Channels, N-Type / drug effects
  • Calcium Channels, N-Type / metabolism*
  • Calcium Signaling / physiology*
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Dose-Response Relationship, Drug
  • Female
  • Heterotrimeric GTP-Binding Proteins / drug effects
  • Heterotrimeric GTP-Binding Proteins / metabolism*
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Models, Biological
  • Molecular Sequence Data
  • Neurons / drug effects
  • Neurons / metabolism*
  • Oocytes
  • Peptide Fragments / pharmacology
  • Protein Binding / physiology
  • Protein Structure, Tertiary / drug effects
  • Protein Structure, Tertiary / physiology
  • Recombinant Fusion Proteins / drug effects
  • Recombinant Fusion Proteins / pharmacology
  • Synapses / drug effects
  • Synapses / metabolism*
  • Synaptic Transmission / drug effects
  • Synaptic Transmission / physiology*
  • Xenopus laevis

Substances

  • Calcium Channels, N-Type
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • voltage-dependent calcium channel (P-Q type)
  • Heterotrimeric GTP-Binding Proteins