Pharmacological and electrophysiological properties of the naturally occurring Pro391Arg variant of the human 5-HT3A receptor.
The 5-HT3A receptor, a ligand-gated ion channel, is involved in pain pathways, nausea and emesis, and irritable bowel syndrome, and may play a role in the pathogenesis of psychiatric diseases such as schizophrenia and depression. Recently, a naturally occurring variation (ProArg) in the second intracellular loop of the human (h) 5-HT3A receptor was identified in a schizophrenic patient. Because the substitution of proline, an alpha-imino acid, by arginine may affect the conformation of the whole receptor, the aim of the present study was to determine the pharmacological and functional properties of this variant compared to the wild-type receptor in stably transfected HEK293 cells. Studies of binding of [H]GR65630, a 5-HT3 receptor antagonist, to membranes (saturation and competition experiments with 5-HT3 receptor ligands) and patch-clamp studies of agonist-induced currents in outside-out patches were carried out. In comparison to the wild-type, the variant receptor exhibited no changes in the receptor density and the affinities for nine representative ligands (five agonists and four antagonists). The potencies and efficacies of three 5-HT3 receptor agonists in inducing currents through the ion channel and the potencies of two 5-HT3 receptor antagonists in blocking 5-HT-evoked currents did not differ between wild-type and variant receptors. In addition, there were no differences in the desensitization kinetics of both receptor isoforms. In conclusion, the ArgPro variation of the h5-HT3A receptor does not change ligand binding to the h5-HT3A receptor, nor does it modify current through the receptor channel.
Kurzwelly
Delia
D
Institute of Pharmacology and Toxicology, University of Bonn, Clinic of Anaesthesiology and Intensive Care Medicine, University Clinics of Bonn, Bonn, Germany.
Barann
Martin
M
Kostanian
Arevat
A
Combrink
Sandra
S
Bönisch
Heinz
H
Göthert
Manfred
M
Brüss
Michael
M
D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism*; D004594:Electrophysiology; D014644:Genetic Variation; D006801:Humans; D007093:Imidazoles / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism*; D007473:Ion Channels / Q000502:physiology; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D007930:Leucine / Q000737:chemistry; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D018408:Patch-Clamp Techniques; D011392:Proline / Q000737:chemistry; D011485:Protein Binding; D011869:Radioligand Assay; D044406:Receptors, Serotonin, 5-HT3 / Q000378:metabolism*; D012701:Serotonin / Q000378:metabolism; D058827:Serotonin 5-HT3 Receptor Agonists; D058831:Serotonin 5-HT3 Receptor Antagonists; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology
D007093:Imidazoles; D007211:Indoles; D007473:Ion Channels; D008024:Ligands; D044406:Receptors, Serotonin, 5-HT3; D058827:Serotonin 5-HT3 Receptor Agonists; D058831:Serotonin 5-HT3 Receptor Antagonists; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; C059068:GR 65630; D012701:Serotonin; D011392:Proline; D007930:Leucine