Abstract
A tyrosine residue at the cytoplasmic end of the seventh transmembrane
helix is conserved in many G-protein-coupled receptors. In the human
beta 2-adrenoceptor, this tyrosine (Tyr326) has been proposed to
be a specific determinant for agonist-induced receptor sequestration.
In order to probe its contribution to the sequestration process we
have replaced this tyrosine by alanine (Y326A) or phenylalanine (Y326F).
Wild-type and mutant receptors were stably expressed in Chinese hamster
ovary cells. Agonist-induced sequestration was essentially abolished
in Y326A receptors and only slightly reduced in Y326F receptors.
However, cells expressing Y326A receptors displayed a high percentage
of internal receptors under basal conditions while cells expressing
wild-type receptors did not. In addition, high-affinity agonist binding
and the ability to activate adenylyl cyclase were markedly reduced
in Y326A receptors and slightly reduced in Y326F receptors. We conclude
that Tyr326 is required for the functional integrity of the beta
2-adrenoceptor and that it may be involved in multiple agonist-induced
effects.
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