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. 2008 Dec 12;417(Pt 1):161–172. doi: 10.1042/BJ20081227

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© 2009 The Author(s) The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Non-Commercial Licence (http://creativecommons.org/licenses/by-nc/2.5/) which permits unrestricted non-commercial use, distribution and reproduction in any medium, provided the original work is properly cited.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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(A) Ligand-binding characteristics of the wild-type and mutant α_1b-adrenoceptor. The capacity of phenylephrine to compete for binding with [³H]prazosin to FLAG-α_1b-adrenoceptor-eYFP (■) and FLAG-α_1b-adrenoceptor TMI-TMIV-eYFP (□) was assessed. (B) Stimulation of [³⁵S]GTP[S] binding by phenylephrine in Gα_q/Gα₁₁ immunoprecipitates from membranes of HEK-293 cells stably expressing FLAG-α_1b-adrenoceptor-eYFP (■) and FLAG-α_1b-adrenoceptor TMI-TMIV-eYFP (□). (C) Intracellular calcium mobilization induced by phenylephrine in cells stably expressing FLAG-α_1b-adrenoceptor-eYFP (■) and FLAG-α_1b-adrenoceptor TMI-TMIV-eYFP previously treated with 10⁻⁷ M prazosin (□). Complete removal of prazosin after the overnight period was assessed by testing the response of the wild-type receptor after treatment and washout (open circles). All graphs are representative results from at least three independent experiments.