New roles for RGS2, 5 and 8 on the ratio-dependent modulation of recombinant GIRK channels expressed in Xenopus oocytes

J Physiol. 1999 Jun 1;517 ( Pt 2)(Pt 2):341-52. doi: 10.1111/j.1469-7793.1999.0341t.x.

Abstract

1. The activation of G protein-regulated inward rectifying potassium (GIRK) channels is modulated by G protein-coupled receptors (GPCRs) via the G protein betagamma subunits and is accelerated by regulators of G protein signalling (RGS). In the present study we investigated the ratio dependence of receptor-mediated activation and deactivation and the influence of new members of the RGS protein family on GIRK currents by coexpressing the recombinant protein subunits in Xenopus oocytes and further analysis of the whole cell currents. 2. The activation of GIRK channels by the muscarinic acetylcholine receptor M2 (M2 mAChR) is strongly dependent on the ratio of receptor to channel in Xenopus oocytes. The increase and on-rate of the amplified current is affected by this ratio. An excess of receptor over channel is necessary for current amplification, while the reverse excess of channel over receptor abolishes the effect. 3. The speed of receptor-mediated activation of GIRK currents is accelerated for a high ratio of receptor to channel, while the time of deactivation is independent of this ratio. 4. Coexpression of RGS2, 5 and 8 accelerates the speed for ACh-mediated activation and deactivation of GIRK1/2 and GIRK1/4 currents. Thereby the receptor/channel/RGS ratio determines the amount of current amplification. 5. Bordetella pertussis toxin completely abolished ACh-mediated current amplification of GIRK channels coexpressed with or without RGS2. 6. Two single point mutations in the RGS2 protein (RGS2(N109S) and RGS2(L180F)) reduced the acceleration of current amplification after ACh application on GIRK1/4 channels compared with RGS2 wild-type protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence / genetics
  • Animals
  • Electric Conductivity
  • Female
  • GTP-Binding Proteins / physiology*
  • GTPase-Activating Proteins
  • Mice
  • Molecular Sequence Data
  • Oocytes / metabolism
  • Oocytes / physiology
  • Pertussis Toxin
  • Point Mutation / physiology
  • Potassium Channels / metabolism
  • Potassium Channels, Inwardly Rectifying*
  • Proteins / genetics
  • Proteins / physiology*
  • RGS Proteins*
  • Rats
  • Receptors, Muscarinic / physiology
  • Recombinant Proteins
  • Virulence Factors, Bordetella / pharmacology
  • Xenopus Proteins*
  • Xenopus laevis / metabolism

Substances

  • GTPase-Activating Proteins
  • Potassium Channels
  • Potassium Channels, Inwardly Rectifying
  • Proteins
  • RGS Proteins
  • Receptors, Muscarinic
  • Recombinant Proteins
  • Rgs2 protein, mouse
  • Rgs5 protein, Xenopus
  • Rgs5 protein, mouse
  • Rgs5 protein, rat
  • Virulence Factors, Bordetella
  • Xenopus Proteins
  • RGS4 protein
  • Pertussis Toxin
  • GTP-Binding Proteins

Associated data

  • GENBANK/AB006013
  • GENBANK/U67187
  • GENBANK/U67188