Steroid and G Protein Binding Characteristics of the Seatrout and Human Progestin Membrane Receptor Alpha Subtypes and Their Evolutionary Origins

A novel progestin receptor (mPR) with seven-transmembrane domains was recently discovered in spotted seatrout and homologous genes were identified in other vertebrates. We show that cDNAs for the mPR α subtypes from spotted seatrout (st-mPRα) and humans (hu-mPRα) encode progestin receptors that display many functional characteristics of G protein-coupled receptors. Flow cytometry and immunocytochemical staining of whole MDA-MB-231 cells stably transfected with the mPRαs using antibodies directed against their N-terminal regions show the receptors are localized on the plasma membrane and suggest the N-terminal domain is extracellular. Both recombinant st-mPRα and hu-mPRα display high affinity (Kd 4.2–7.8 nm), limited capacity (Bmax 0.03–0.32 nm), and displaceable membrane binding specific for progestins. Progestins activate a pertussis toxin-sensitive inhibitory G protein (Gi) to down-regulate membrane-bound adenylyl cyclase activity in both st-mPRα- and hu-mPRα-transfected cells. Coimmunoprecipitation exp...