Adenosine receptor permanently coupled to turkey erythrocyte adenylate cyclase

Abstract

The mode of coupling of the adenosine receptor to adenylate cyclase in turkey erythrocyte membranes was probed by 2 independent approaches. The progressive inactivation of the adenosine receptor by an adenosine receptor affinity label resulted in the proportional reduction in the adenosine plus GppNHp dependent specific activity. The intrinsic rate constant (k3), characterizing the process of adneylate cyclase activation by the adenosine-adenosine receptor complex, is independent of the extent of receptor inactivation. This behavior favored the precoupled mechanism, A + R.cntdot.E .**GRAPHIC**. A.cntdot.R.cntdot.E .**GRAPHIC**. ARE'', where the receptor R and the enzyme E are permanently coupled to each other and the adenosine A binds to the receptor and induces the 1st-order process of cyclase activation to its active form ARE'', The finding that adenosine receptor was permanently coupled to the cyclase catalytic unit was corroborated by the observation that the progressive increase in membrane fluidity had no effect on the rate constant (k3) of adenylate cyclase activation by the adenosine-adenosine receptor complex and that the dose-response curve for adenosine was noncooperative.