title: Identification of receptor domains that modify ligand binding to 5-hydroxytryptamine2 and 5-hydroxytryptamine1c serotonin receptors.
abstract: Serotonin [5-hydroxytryptamine (5-HT)] receptors are distinguished pharmacologically by their characteristic affinities for agonists and antagonists. Two serotonin receptors, the 5-HT2 and 5-HT1c, share a number of pharmacologic and structural properties while differing in their affinities for certain agonists and antagonists. To identify regions of the 5-HT2 and 5-HT1c receptors important for specifying their unique pharmacology, we constructed six chimeric 5-HT2/5-HT1c receptors in which domains of each receptor were exchanged. The abilities of several drugs to inhibit [3H]mesulergine bound to the chimeric and parent receptors transiently expressed in COS-7 cells were then examined. For spiperone and haloperidol (both butyrophenones), chimeras that exchanged transmembrane (TM) domains I and II or TMs I-III had the greatest effects on binding affinity. The binding affinity of cinanserin (a cinnamanilide) was significantly changed in all the chimeras studied. In contrast, the binding of ketanserin (a 4-fluorobenzoylpiperidine) was strongly influenced by chimeras that exchanged TMs I-III (but not I and II) and by chimeras that exchanged intracellular loop 3 to TM VII. 5-HT binding affinity was greatly altered for chimeras that exchanged domains of intracellular loop 3 to TM VII, with minor effects being noted for chimeras that exchanged TMs I and II and I-III. The affinities of the nonselective drugs mesulergine, mianserin, and m-chlorophenylpiperazine were relatively unaffected when domains of the 5-HT2 and 5-HT1c receptors were exchanged. Taken together, these results imply that structurally diverse 5-HT2 antagonists utilize distinct regions of the 5-HT2 receptor for high affinity binding.
authors.affiliation: Department of Psychiatry, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106.
authors.forename: M S
authors.initials: MS
authors.lastname: Choudhary
authors.forename: S
authors.initials: S
authors.lastname: Craigo
authors.forename: B L
authors.initials: BL
authors.lastname: Roth
mesh_terms: D001483:Base Sequence; D001667:Binding, Competitive; D004873:Ergolines / Q000378:metabolism; D006220:Haloperidol / Q000378:metabolism; D066298:In Vitro Techniques; D007650:Ketanserin / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011985:Receptors, Serotonin / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013134:Spiperone / Q000378:metabolism; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D004873:Ergolines; D009838:Oligodeoxyribonucleotides; D011985:Receptors, Serotonin; D011993:Recombinant Fusion Proteins; D013134:Spiperone; D007650:Ketanserin; D006220:Haloperidol; C031649:mesulergine