title: Two human alpha 2-adrenoceptor subtypes alpha 2A-C10 and alpha 2B-C2 expressed in Sf9 cells couple to transduction pathway resulting in opposite effects on cAMP production.
abstract: The baculovirus expression vector system utilizing the strong polyhedrin gene promoter of the Autographa californica nuclear polyhedrosis virus (AcNPV) was used for high level expression of the two alpha 2-adrenoceptor subtypes alpha 2A-C10 and alpha 2B-C2 in Spodoptera frugiperda (Sf-9) insect cells. For rapid screening of recombinant viruses the luciferase gene was expressed under the early ETL-promoter (early transcript large) in the same plasmid. Both receptor subtypes showed the same rank order of binding affinity for four agonists tested: dexmedetomidine > l-medetomidine = clonidine > noradrenaline. For the alpha 2A-C10 subtype, these agonists inhibited forskolin stimulated cAMP production through pertussis toxin sensitive G-proteins. In contrast, for the alpha 2B-C2 subtype the agonists stimulated both basal and forskolin stimulated cAMP production.
authors.affiliation: Department of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland.
authors.forename: C C
authors.initials: CC
authors.lastname: Jansson
authors.forename: M
authors.initials: M
authors.lastname: Karp
authors.forename: C
authors.initials: C
authors.lastname: Oker-Blom
authors.forename: J
authors.initials: J
authors.lastname: Näsman
authors.forename: J M
authors.initials: JM
authors.lastname: Savola
authors.forename: K E
authors.initials: KE
authors.lastname: Akerman
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism*; D004274:DNA, Recombinant / Q000302:isolation & purification; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D008156:Luciferases / Q000235:genetics; D010957:Plasmids / Q000502:physiology; D011869:Radioligand Assay; D018341:Receptors, Adrenergic, alpha-2 / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera; D014161:Transduction, Genetic
chemical_list: D004274:DNA, Recombinant; D018341:Receptors, Adrenergic, alpha-2; D000242:Cyclic AMP; D008156:Luciferases