Threonine-for-leucine mutation within domain M2 of the neuronal α 7 nicotinic receptor converts 5-hydroxytryptamine from antagonist to agonist

E. Palma, A. M. Mileo, F. Eusebi, R. Miledi

Research output: Contribution to journalArticlepeer-review

Abstract

A study was made of the effects of 5-hydroxy-tryptamine (5HT) on homomeric neuronal nicotinic receptors (nAcChoR) expressed in Xenopus oocytes after injection of cDNA encoding the wild-type chicken α 7 subunit. Acetylcholine (AcCho) elicited large currents (I(AcChu)) that were reduced by 5HT in a reversible and dose-dependent manner, with a half-inhibitory concentration (IC 50) of 56 μM and a Hill coefficient (nH) of 1.2. The inhibition of IAacCHO BY %HT by 5HT was noncompetitive and voltage independent, a behavior incompatible with a channel blockade mechanism. 5HT alone did not elicit membrane currents in oocytes injected with the wild- type α 7 subunit cDNA. In contrast, 5HT elicited membrane currents (I5(HT)) in oocytes injected with cDNA encoding an α 7 mutant subunit with a threonine-for-leucine-247 substitution (L247T α 7). I(5HT) was inhibited by the potent nicotinic receptor blockers α-bungarotoxin (100 nM) and methyllycaconitine (1 μM). Furthermore, the characteristics of I(5HT), including its voltage dependence, were similar to those of I(AcCho). The 5HT dose-I(5HT) response gave an apparent dissociation constant EC 50 of 23.5 μM and a Hill coefficient n(H) of 1.7, which were not modified by the presence of AcCho. Similarly, the apparent affinity of L247T α 7 for AcCho as well as its cooperativity were not influenced by 5HT, indicating a lack of mutual interactions between 5HT and AcCho. These results show that 5HT is a potent noncompetitive antagonist of neuronal α 7 nAcChoR, but it becomes a noncompetitive agonist following mutation of the highly conserved leucine residue 247 located in the channel domain M2.

Original languageEnglish
Pages (from-to)11231-11235
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume93
Issue number20
DOIs
Publication statusPublished - Oct 1 1996

Keywords

  • acetylcholine receptors
  • neurotransmitter interactions
  • Xenopus oocytes

ASJC Scopus subject areas

  • Genetics
  • General

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