Selective photoinduced uncoupling of the response of adenylate cyclase to gonadotropins by 5-iodonaphthyl 1-azide.
5-Iodonaphthyl 1-azide (INA) has been previously shown to selectively label, on photolysis, only those proteins in contact with the membrane lipids. Low concentrations (less than 10 microM) of INA added to rat ovarian plasma membranes induced, on photoactivation, a selective and complete loss of the response of the adenylate cyclase to stimulation by human chorionic gonadotropin (hCG) or luteinizing hormone (LH). In contrast, this treatment affected neither hCG binding to the receptor nor the stimulation of the enzyme by NaF. That the uncoupling of the receptor from the enzyme by INA occurred within the lipid bilayer can be derived from the finding that the prior presence neither of saturating concentrations of hCG nor of the aqueous nitrene-scavenger glutathione (GSH) prevented this effect. Photolysis at higher concentrations of INA (0.1-1 mM) led to the inhibition of the adenylate cyclase stimulated by fluoride. This effect was totally prevented by glutathione. A similar behavior was obtained with a water-soluble analogue of INA, namely, 5-diazonionapthyl 1-azide (DAN). On photoactivation with 30 microM DAN, the NaF-stimulated adenylate cyclase was inhibited, but this effect was completely prevented by added GSH. At low concentrations where its effects are restricted to the lipid core, INA may represent a useful tool to define receptor coupling with the adenylate cyclase. The capacity of INA at low concentrations to uncouple the hormone receptor from the adenylate cyclase is not restricted to the LH/hCG receptor. Other hormone receptors tested behaved similarly. Therefore, the reported findings appear to represent a general phenomenon.
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D001386:Azides / Q000494:pharmacology*; D002462:Cell Membrane / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000378:metabolism / Q000494:pharmacology*; D004789:Enzyme Activation; D005260:Female; D007457:Iodine Radioisotopes; D007700:Kinetics; D007986:Luteinizing Hormone / Q000378:metabolism / Q000494:pharmacology*; D010053:Ovary / Q000378:metabolism; D010782:Photolysis; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D011974:Receptors, LH
D001386:Azides; D006063:Chorionic Gonadotropin; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D011974:Receptors, LH; C015298:iodonaphthylazide; D007986:Luteinizing Hormone; D000262:Adenylyl Cyclases