Role of the extracellular transmembrane domain interface in gating and pharmacology of a heteromeric neuronal nicotinic receptor.
Nicotinic acetylcholine receptors (nAChRs) transmit the agonist signal to the channel gate through a number of extracellular domains. We have previously shown that particular details of the process of coupling binding to gating could be quantitative and qualitatively different in muscle and neuronal type nAChRs. We have extended previous studies on homomeric alpha7 nAChRs to heteromeric alpha3beta4 nAChRs, by mutating residues located at loops 2 and 7, and M2-M3 linker of both alpha3 and beta4 subunits which, in order to monitor surface expression, were modified to bind alpha-bungarotoxin, and expressed in Xenopus oocytes. We show that, in general, mutations in these domains of both alpha3 and beta4 subunits affect the gating function, although the effects are slightly larger if they are inserted in the alpha3 subunit. However, the involvement of a previously reported intrasubunit interaction in coupling (Gln48-Ile130) seems to be restricted to the beta4 subunit. We also show that mutations at these domains, particularly loop 2 of the alpha3 subunit, change the pharmacological profile of alpha3beta4 nAChRs, decreasing nicotine's and increasing cytisine's effectiveness relative to acetylcholine. It is concluded that, unlike muscle nAChRs, the non-alpha subunits play a relevant role in the coupling process of neuronal alpha3beta4 nAChRs.
Aldea
Marcos
M
Instituto de Neurociencias de Alicante, Universidad Miguel Hernández-CSIC, Sant Joan d'Alacant, Alicante, Spain.
Castillo
Mar
M
Mulet
José
J
Sala
Salvador
S
Criado
Manuel
M
Sala
Francisco
F
D000109:Acetylcholine / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D005260:Female; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000187:drug effects / Q000235:genetics; D009865:Oocytes; D017434:Protein Structure, Tertiary / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D021122:Protein Subunits / Q000737:chemistry / Q000187:drug effects / Q000235:genetics; D011978:Receptors, Nicotinic / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics; D009435:Synaptic Transmission / Q000187:drug effects / Q000235:genetics; D014982:Xenopus laevis
D000109:Acetylcholine; D009419:Nerve Tissue Proteins; D021122:Protein Subunits; D011978:Receptors, Nicotinic; C447159:CHRNB4 protein, human