title: Nicotine enhances the cyclic AMP-dependent protein kinase-mediated phosphorylation of alpha4 subunits of neuronal nicotinic receptors.
abstract: Studies determined whether alpha4beta2 or alpha3beta2 neuronal nicotinic receptors expressed in Xenopus oocytes are substrates for cyclic AMP-dependent protein kinase (PKA) and whether nicotine affects receptor phosphorylation. The cRNAs for the subunits were coinjected into oocytes, and cells were incubated for 24 h in the absence or presence of nicotine (50 nM for alpha4beta2 and 500 nM for alpha3beta2 receptors). Nicotine did not interfere with the isolation of the receptors. When receptors isolated from oocytes expressing alpha4beta2 receptors were incubated with [gamma-32P]ATP and the catalytic subunit of PKA, separated by electrophoresis, and visualized by autoradiography, a labeled phosphoprotein with the predicted molecular size of the alpha4 subunit was present. Phosphorylation of alpha4 subunits of alpha4beta2 receptors increased within the first 5 min of incubation with nicotine and persisted for 24 h. In contrast, receptors isolated from oocytes expressing alpha3beta2 receptors did not exhibit a labeled phosphoprotein corresponding to the size of the alpha3 subunit. Results suggest that the PKA-mediated phosphorylation of alpha4 and not alpha3 subunits may explain the differential inactivation by nicotine of these receptor subtypes expressed in oocytes.
authors.affiliation: Department of Pharmacology and Therapeutics, University of South Florida College of Medicine, Tampa 33612-4799, U.S.A.
authors.forename: Y N
authors.initials: YN
authors.lastname: Hsu
authors.forename: S C
authors.initials: SC
authors.lastname: Edwards
authors.forename: L
authors.initials: L
authors.lastname: Wecker
mesh_terms: D000818:Animals; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D009474:Neurons / Q000378:metabolism*; D009538:Nicotine / Q000494:pharmacology*; D009865:Oocytes / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D011978:Receptors, Nicotinic / Q000378:metabolism*; D014982:Xenopus laevis
chemical_list: D011978:Receptors, Nicotinic; D009538:Nicotine; D017868:Cyclic AMP-Dependent Protein Kinases