Cloning and functional expression of a human 5-hydroxytryptamine type 3AS receptor subunit.
A human 5-hydroxytryptamine receptor type 3AS (5-HT3R-AS) subunit has been cloned from an amygdala cDNA library. We report the nucleotide and predicted amino acid sequence of the human subunit, which possesses 85% and 84% amino acid sequence identity with mouse and rat 5-HT3R-AS subunits, respectively. Acting on Xenopus laevis oocytes injected with RNA transcripts of the clone, 5-HT and selective 5-HT3 receptor agonists elicited inwardly directed current responses that displayed desensitization. Such currents were blocked in a concentration-dependent manner by selective and nonselective 5-HT3 receptor antagonists but were unaffected by compounds acting at G protein-linked 5-HT receptors. A quantitative comparison of the pharmacological profiles of human and mouse recombinant 5-HT3R-AS receptor complexes revealed differences in the potencies of some antagonist or agonist compounds tested, the most dramatic example being (+)-tubocurarine, which demonstrated an approximately 1800-fold discrepancy in antagonist potency. In view of the small number of sequence substitutions that occur between the human and mouse homologues of the 5-HT3R-AS in the extracellularly located aminoterminal domain, compounds such as (+)-tubocurarine, in conjunction with site-directed mutagenesis, may prove to be valuable in locating amino acid residues that contribute to the ligand binding site(s) of the 5-HT3 receptor. Also, when methodological differences are taken into account, the present study suggests that a homo-oligomeric assembly of human 5-HT3R-AS subunits can account for the distinctive ligand binding properties of human 5-HT3 receptors established in postmortem brain tissue.
Belelli
D
D
Department of Pharmacology and Clinical Pharmacology, Ninewells Hospital and Medical School, The University, Dundee, UK.
Balcarek
J M
JM
Hope
A G
AG
Peters
J A
JA
Lambert
J J
JJ
Blackburn
T P
TP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004594:Electrophysiology; D006168:Guinea Pigs; D006801:Humans; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes / Q000648:ultrastructure; D011817:Rabbits; D051381:Rats; D011985:Receptors, Serotonin / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D014982:Xenopus laevis
D046911:Macromolecular Substances; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists