A single point mutation decreases picrotoxinin sensitivity of the human GABA receptor rho 1 subunit.
The GABA receptor rho subunits are thought to form bicuculline-insensitive and picrotoxinin-sensitive GABAC receptors. We have investigated the role of the amino acid at position 309 in transmembrane segment M2 of the human rho 1 subunit as a determinant for picrotoxinin sensitivity. The mutant rho 1P309S was constructed by exchanging proline 309 for serine, the corresponding amino acid of the human rho 2 subunit. Whole-cell recordings from HEK-293 cells transfected with rho 1P309S cDNA revealed that the sensitivity of the rho 1P309S channels for picrotoxinin was four-fold lower than that of the wild type rho 1 subunit. The affinity of the mutant receptor for GABA was only slightly changed. These results provide direct evidence that the amino acid at position 309 is an important determinant for the picrotoxinin sensitivity of GABA receptors formed by the rho subunits.
Enz
R
R
Max-Planck Institut für Hirnforschung, Frankfurt, Germany.
Bormann
J
J
D001483:Base Sequence; D001640:Bicuculline / Q000494:pharmacology; D002460:Cell Line; D006801:Humans; D008969:Molecular Sequence Data; D018408:Patch-Clamp Techniques; D010446:Peptide Fragments / Q000737:chemistry / Q000187:drug effects*; D010852:Picrotoxin / Q000031:analogs & derivatives* / Q000494:pharmacology; D017354:Point Mutation; D011392:Proline / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D018079:Receptors, GABA / Q000737:chemistry / Q000187:drug effects*; D012694:Serine / Q000737:chemistry*; D054830:Sesterterpenes
D001640:Bicuculline; D010446:Peptide Fragments; D010852:Picrotoxin; D011392:Proline; D018079:Receptors, GABA; D012694:Serine; D054830:Sesterterpenes; C022961:picrotoxinin