Identification of an amino acid defining the distinct properties of murine beta1 and beta3 subunit-containing GABA(A) receptors.
Murine gamma-aminobutyric acid (GABA) type A homomeric receptors made of beta1 subunits are profoundly different, when expressed in Xenopus oocytes, from beta3 homomeric receptors. Application of the intravenous general anesthetic pentobarbital, etomidate, or propofol to beta3 homomeric receptors allows current flow. In contrast, beta1 homomers do not respond to any of these agents. Through construction of chimeric beta1/beta3 receptors, we identified a single amino acid that determines the pharmacological difference between the two beta subunits. When the serine residue present in the wild-type nonresponsive beta1 subunit is replaced by an asparagine found in the same position in the beta3 subunit, the resulting point-mutated beta1S265N forms receptors responsive to intravenous general anesthetics, like the wild-type beta3 subunits. Conversely, after mutation of the wild-type beta3 to beta3N265S, the homomeric receptor loses its ability to respond to these same general anesthetics. Wild-type-to-mutant titration experiments showed that the nonresponsive phenotype is dominant: A single nonresponsive residue within a pentameric receptor is sufficient to render the receptor nonresponsive. In alpha1betax or alpha1betaxgamma2 heteromeric receptors, the same residue manifests as a partial determinant of the degree of potentiation of the GABA-induced current by some general anesthetics. The location of this amino acid at the extracellular end of the second transmembrane segment, its influence in both homomeric and heteromeric receptor function, and its dominant behavior suggest that this residue of the beta subunit is involved in an allosteric modulation of the receptor.
Cestari
I N
IN
Neuroscience Graduate Program, University of Texas Southwestern Medical Center, Dallas, USA.
Min
K T
KT
Kulli
J C
JC
Yang
J
J
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes; D010424:Pentobarbital / Q000494:pharmacology; D020033:Protein Isoforms / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011963:Receptors, GABA-A / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D012694:Serine / Q000235:genetics; D014982:Xenopus laevis
D007473:Ion Channels; D020033:Protein Isoforms; D011963:Receptors, GABA-A; D012694:Serine; D010424:Pentobarbital