Mutations in the channel domain alter desensitization of a neuronal nicotinic receptor.
A variety of ligand-gated ion channels undergo a fast activation process after the rapid application of agonist and also a slower transition towards desensitized or inactivated closed channel states when exposure to agonist is prolonged. Desensitization involves at least two distinct closed states in the acetylcholine receptor, each with an affinity for agonists higher than those of the resting or active conformations. Here we investigate how structural elements could be involved in the desensitization of the acetylcholine-gated ion channel from the chick brain alpha-bungarotoxin sensitive homo-oligomeric alpha 7 receptor, using site-directed mutagenesis and expression in Xenopus oocytes. Mutations of the highly conserved leucine 247 residue from the uncharged MII segment of alpha 7 suppress inhibition by the open-channel blocker QX-222, indicating that this residue, like others from MII, faces the lumen of the channel. But, unexpectedly, the same mutations decrease the rate of desensitization of the response, increase the apparent affinity for acetylcholine and abolish current rectification. Moreover, unlike wild-type alpha 7, which has channels with a single conductance level, the leucine-to-threonine mutant has an additional conducting state active at low acetylcholine concentrations. It is possible that mutation of Leu 247 renders conductive one of the high-affinity desensitized states of the receptor.
Revah
F
F
Neurobiologie Moléculaire, Unité de Recherche Associée au Centre National de la Recherche Scientifique, Institut Pasteur, Paris, France.
Bertrand
D
D
Galzi
J L
JL
Devillers-Thiéry
A
A
Mulle
C
C
Hussy
N
N
Bertrand
S
S
Ballivet
M
M
Changeux
J P
JP
D000109:Acetylcholine / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000502:physiology*; D002645:Chickens; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009474:Neurons / Q000502:physiology*; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D011978:Receptors, Nicotinic / Q000235:genetics / Q000502:physiology*; D014981:Xenopus
D007473:Ion Channels; D011978:Receptors, Nicotinic; D000109:Acetylcholine