title: Domain near TM1 influences agonist and antagonist responses of peptide-gated Na+ channels.
abstract: A molecular biological approach was used to analyse the importance of different amino acids for ligand activation and for determining the action of amiloride on peptide- (Phe-Met-Arg-Phe-NH(2), FMRFamide)-gated Na(+) channels, members of the degenerin/epithelial Na channel (DEG/ENaC) family. Amiloride is a low-affinity blocker of most DEG/ENa channels, but has an unusual enhancing effect on responses of some of them. Chimeras were expressed in Xenopus oocytes and studied electrophysiologically. Differences in properties of channels from Helix aspersa and Helisoma trivolvis highlighted a sequence of 50 residues of the extracellular domain, near the first transmembrane segment (TM1), that affected sensitivity to FMRFamide, and whether amiloride blocked or enhanced the response to FMRFamide. Comparisons of chimeras prepared from H. aspersa and the extracellular domains of two other species, Aplysia californica and Lymnaea stagnalis and the preparation of further constructs, showed that amino acids 128-134 in the H. aspersa sequence are important in determining the predominant effect of amiloride and influencing the EC(50) of FMRFamide. The results also showed that amino acids in this region are influenced by amino acids in other regions of the extracellular domain so as to affect not only the magnitude of responses, but also their time course and desensitisation.
authors.affiliation: School of Biology, University of St Andrews, Fife, KY16 9TS, UK. gac@st-and.ac.uk
authors.forename: Glen A
authors.initials: GA
authors.lastname: Cottrell
mesh_terms: D000584:Amiloride / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001048:Aplysia / Q000235:genetics; D002678:Chimera / Q000235:genetics; D019835:FMRFamide / Q000819:agonists / Q000037:antagonists & inhibitors; D006372:Helix, Snails / Q000235:genetics; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D008195:Lymnaea / Q000235:genetics; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000502:physiology*; D012908:Snails / Q000235:genetics; D015222:Sodium Channels / Q000235:genetics* / Q000502:physiology*; D014981:Xenopus
chemical_list: C097095:FaNaCh protein, Helix aspersa; D015222:Sodium Channels; D019835:FMRFamide; D000584:Amiloride