Downregulation of bradykinin B2 receptor in human fibroblasts during prolonged agonist exposure

• Sustained activation of G protein-coupled receptors results in an attenuation of cellular responses, a phenomenon termed desensitization. Whereas mechanisms for rapid desensitization of ligand-receptor-G protein-effector systems are relatively well characterized, much less is known about long-term adaptation processes that occur in the continuous presence of an agonist. Here we have studied the fate of endogenously expressed bradykinin B(2) receptors on human fibroblasts during prolonged agonist treatment. Stimulation with bradykinin for up to 24 h resulted in a 50% reduction of surface binding sites that was paralleled by a similar decrease of total B(2) receptor protein followed by Western blotting using monoclonal antibodies to the B(2) receptor. Whereas B(2) receptor mRNA levels did not change during 24 h of agonist treatment, B(2) receptor de novo synthesis was attenuated by 35-50%, indicating translational control of B(2) receptor levels. Furthermore, the half-life of B(2) receptor protein was shortened by 20-40% as shown by (35)S-labeled pulse-chase and immunoprecipitation experiments. This study demonstrates that bradykinin B(2) receptor expression during long-term agonist treatment is primarily regulated on the (post)translational level, i.e., by attenuation of de novo synthesis and by reduction of receptor stability.

Nyckelord

Antibodies; Monoclonal/diagnostic use

Blotting; Northern

Blotting; Western

Bradykinin/pharmacology

Cells; Cultured

Down-Regulation/*drug effects

Fibroblasts/metabolism

Half-Life

Humans

Protein Processing; Post-Translational/physiology

RNA; Messenger/biosynthesis/genetics

Radioligand Assay

Receptor; Bradykinin B2

Receptors; Bradykinin/*agonists

Stimulation; Chemical

Transcription; Genetic/drug effects

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)