Identification of an amino acid sequence within GABA(A) receptor beta3 subunits that is important for receptor assembly.
GABA(A) receptors are chloride ion channels that can be opened by GABA, the most important inhibitory transmitter in the CNS. In the mammalian brain the majority of these pentameric receptors is composed of two alpha, two beta and one gamma subunit. To achieve the correct order of subunits around the pore, each subunit must form specific contacts via its plus (+) and minus (-) side. To identify a sequence on the beta3 subunit important for assembly, we generated various full-length or truncated chimeric beta3 constructs and investigated their ability to assemble with alpha1 and gamma2 subunits. It was demonstrated that replacement of the sequence beta3(76-89) by the homologous alpha1 sequence impaired assembly with alpha1 but not with gamma2 subunits in alpha1beta3gamma2-GABA(A) receptors. Other experiments indicated that assembly was impaired via the beta3(-) side of the chimeric subunit. Within the sequence beta3(76-89) the sequence beta3(85-89) seemed to be of primary importance for assembly with alpha1 subunits. A comparison with the structure of the acetylcholine-binding protein supports the conclusion that the sequence beta3(85-89) is located at the beta3(-) side and indicates that it contains amino acid residues that might directly interact with the (+) side of the neighbouring alpha1 subunit.
Ehya
Noosha
N
Division of Biochemistry and Molecular Biology, Brain Research Institute, University of Vienna, Austria.
Sarto
Isabella
I
Wabnegger
Leila
L
Sieghart
Werner
W
D000595:Amino Acid Sequence / Q000235:genetics; D001665:Binding Sites / Q000502:physiology; D002460:Cell Line; D002678:Chimera; D006801:Humans; D009118:Muscimol / Q000378:metabolism; D020033:Protein Isoforms / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D011963:Receptors, GABA-A / Q000502:physiology*
D020033:Protein Isoforms; D011963:Receptors, GABA-A; D009118:Muscimol