Modulation of 5-HT(1A) receptor activation by its interaction with wild-type and mutant g(alphai3) proteins

Neuropharmacology. 2001;40(1):36-47. doi: 10.1016/s0028-3908(00)00098-8.


Constitutive and agonist-dependent activation of the recombinant human 5-HT(1A) receptor (RC: 2.1.5HT.01A) was investigated by co-expression with a rat G(alphai3) protein in Cos-7 cells. The interaction between the 5-HT(1A) receptor and rat G(alphai3) protein was modulated by substitution of the G(alphai3) protein site for pertussis toxin-catalysed ADP-ribosylation (cysteine(351)) by each of the natural amino acids. Enhanced basal [(35)S]GTPgammaS binding responses (+24 to +189%) were observed with the mutant G(alphai3) proteins containing at position 351 either a histidine, glutamine, serine, tyrosine or a nonpolar amino acid with the exception of a proline. With each of these mutant G(alphai3) proteins, spiperone (10 microM), but not WAY 100635 (10 microM), reduced (-22 to -60%, p<0.05) the enhanced basal [(35)S]GTPgammaS binding response. 5-HT (10 microM)-mediated [(35)S]GTPgammaS binding responses attained for some of the mutant G(alphai3)Cys(351) proteins (Phe, Met, Val and Ala) more than 300% of that obtained with the wt G(alphai3) protein. Similar results were also obtained with the prototypical 5-HT(1A) agonist 8-OH-DPAT and the partial agonist (-)-pindolol. Fusion proteins assembled from the 5-HT(1A) receptor and either the wt G(alphai3)Cys(351), mutant G(alphai3)Cys(351)Gly or G(alphai3)Cys(351)Ile protein displayed similar observations for these ligands as obtained by co-expression of the 5-HT(1A) receptor with each of these G(alphai3) proteins. Both the degree of 5-HT(1A) receptor activation by 8-OH-DPAT and (-)-pindolol, and its inhibition by spiperone, strongly correlate (r(2): 0.78-0.81) with the octanol/water partition coefficients of the mutated amino acid at position 351 of the G(alphai3) protein. The present data also suggest the wt G(alphai3) protein does not result in maximal activation of the 5-HT(1A) receptor by the agonists being investigated.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism
  • Cloning, Molecular
  • GTP-Binding Protein alpha Subunits, Gi-Go / drug effects*
  • GTP-Binding Protein alpha Subunits, Gi-Go / genetics*
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Immunohistochemistry
  • Molecular Sequence Data
  • Mutation / genetics
  • Pertussis Toxin
  • Radioligand Assay
  • Rats
  • Receptors, Serotonin / drug effects*
  • Receptors, Serotonin, 5-HT1
  • Serotonin Receptor Agonists / pharmacology*
  • Structure-Activity Relationship
  • Transfection
  • Virulence Factors, Bordetella / pharmacology


  • Receptors, Serotonin
  • Receptors, Serotonin, 5-HT1
  • Serotonin Receptor Agonists
  • Virulence Factors, Bordetella
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Pertussis Toxin
  • GTP-Binding Protein alpha Subunits, Gi-Go