Receptor-mediated modulation of recombinant neuronal class E calcium channels

FEBS Lett. 1997 May 26;408(3):261-70. doi: 10.1016/s0014-5793(97)00437-7.

Abstract

The modulation of a cloned neuronal calcium channel was studied in a human embryonic kidney cell line (HEK293). The HEK293 cells were stably transfected with the alpha1Ed cDNA, containing the pore forming subunit of a neuronal class E calcium channel. Inward currents of 25 +/- 1.9 pA/pF (n = 79) were measured with the cloned alpha1Ed-subunit. The application of the peptide hormone somatostatin, carbachol, ATP or adenosine reduced the amplitude of Ca2+ and Ba2+ inward currents and exhibited a slowing of inactivation. This inhibitory effect by somatostatin was significantly impaired after pre-incubating the transfected cell line with pertussis toxin (PTX). Internal perfusion of the cells with the G-protein-inactivating agent GDP-beta-S or with the permanently activating agent GTP-gamma-S also attenuated the somatostatin effect. The inhibition indicates that modulation of the alpha1Ed-mediated Ca2+ current involves pertussis toxin-sensitive G-proteins. The block of Ca2+ and Ba2+ inward currents by somatostatin is also found in cells expressing a truncated alpha1Ed-subunit which lacks a 129-bp fragment in the C-terminus. This fragment corresponds to the major structural difference between two native human alpha1E splice variants. As somatostatin inhibits inward currents through both, the cloned alpha1Ed- and the truncated alpha1Ed-DEL-subunit, the hormone-mediated modulation is independent from the presence of the 129-bp insertion in the C-terminus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / pharmacology
  • Adenosine Triphosphate / pharmacology
  • Amino Acid Sequence
  • Barium / metabolism
  • Binding Sites
  • Calcium / metabolism*
  • Calcium Channels / chemistry
  • Calcium Channels, L-Type
  • Carbachol / pharmacology
  • Cell Line
  • Cloning, Molecular
  • GTP-Binding Proteins / physiology
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Guanosine Diphosphate / analogs & derivatives
  • Guanosine Diphosphate / pharmacology
  • Humans
  • Kidney
  • Kinetics
  • Macromolecular Substances
  • Membrane Potentials / drug effects
  • Molecular Sequence Data
  • Neurons / metabolism*
  • Pertussis Toxin
  • Recombinant Fusion Proteins / metabolism
  • Sequence Deletion
  • Sequence Homology, Amino Acid
  • Somatostatin / pharmacology
  • Thionucleotides / pharmacology
  • Transfection
  • Virulence Factors, Bordetella / pharmacology

Substances

  • Calcium Channels
  • Calcium Channels, L-Type
  • Macromolecular Substances
  • Recombinant Fusion Proteins
  • Thionucleotides
  • Virulence Factors, Bordetella
  • Guanosine Diphosphate
  • Barium
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Somatostatin
  • guanosine 5'-O-(2-thiodiphosphate)
  • Adenosine Triphosphate
  • Carbachol
  • Pertussis Toxin
  • GTP-Binding Proteins
  • Adenosine
  • Calcium

Associated data

  • GENBANK/L27745