A combination of human alpha 1 and beta 1 subunits is required for formation of detectable GABA-activated chloride channels in Sf9 cells

Proc Biol Sci. 1992 Dec 22;250(1329):307-12. doi: 10.1098/rspb.1992.0163.

Abstract

The baculovirus expression system was used to produce alpha 1 and beta 1 subunits of the human GABAA receptor in Sf9 cells. In cells infected with both alpha 1 and beta 1 recombinant viruses, GABA elicited an outwardly rectifying chloride current that was blocked by bicuculline and potentiated by pentobarbitone. GABA did not produce detectable currents in cells infected with either alpha 1 or beta 1 recombinant viruses alone. In these cells, and in control (non-infected) Sf9 cells, pentobarbitone depressed the leakage current (Ki = 55 microM). Fluorescently labelled monoclonal antibodies to the alpha 1 subunit showed greater amounts of the alpha 1 subunit in cells infected with only the alpha 1 recombinant virus than in cells co-infected with the alpha 1 and beta 1 recombinant viruses. Fluorescence of the plasma membrane was seen in cells co-infected with the alpha 1 and beta 1 recombinant viruses, but was absent in cells infected with only the alpha 1 recombinant virus. It was concluded that the alpha 1 subunit normally interacts with the beta 1 subunit to be transported to the plasma membrane in Sf9 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cell Line
  • Chloride Channels
  • Electric Conductivity / drug effects
  • Fluorescent Antibody Technique
  • Humans
  • Insecta
  • Ion Channels / drug effects
  • Ion Channels / genetics
  • Ion Channels / physiology*
  • Macromolecular Substances
  • Membrane Potentials / drug effects
  • Membrane Proteins / drug effects
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Microscopy, Fluorescence
  • Recombinant Proteins / drug effects
  • Recombinant Proteins / metabolism
  • Restriction Mapping
  • Transfection
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism
  • gamma-Aminobutyric Acid / pharmacology*

Substances

  • Chloride Channels
  • Ion Channels
  • Macromolecular Substances
  • Membrane Proteins
  • Recombinant Proteins
  • gamma-Aminobutyric Acid
  • beta-Galactosidase