title: The composition of the beta-2 adrenergic receptor oligomer affects its membrane trafficking after ligand-induced endocytosis.
abstract: The beta-2 adrenergic receptor (B2AR) is well known to form oligomeric complexes in vivo, but the functional significance of this process is not fully understood. The present results identify an effect of oligomerization of the human B2AR on the membrane trafficking of receptors after agonist-induced endocytosis in stably transfected human embryonic kidney 293 cells. A sequence present in the cytoplasmic tail of the B2AR has been shown previously to be required for efficient recycling of internalized receptors. Mutation of this sequence was observed to inhibit recycling not only of the receptor containing the mutation but also of the coexpressed wild-type B2AR. Coexpression of recycling-defective mutant B2ARs also enhanced proteolytic degradation of the wild-type B2AR after agonist-induced endocytosis, consistent with trafficking of both receptors to lysosomes in an oligomeric complex. Coexpression of the delta opioid receptor (DOR) at similar levels produced a much smaller effect on endocytic trafficking of the B2AR, even though DOR traverses a similar membrane pathway as recycling-defective mutant B2ARs. Biochemical studies confirmed that B2AR/B2AR-ala homomeric complexes form more readily than DOR/B2AR heteromers in expression-matched cell clones and support the hypothesis that B2AR/B2AR-ala complexes are not disrupted by agonist. These results suggest that a significant fraction of B2ARs exists in oligomeric complexes after ligand-induced endocytosis and that the composition of the oligomeric complex influences the sorting of endocytosed receptors between functionally distinct recycling and degradative membrane pathways.
authors.affiliation: Genentech Hall, Rm N212E, University of California-San Francisco, 600 16th Street, San Francisco, CA 94143-2140, USA.
authors.forename: Tracy T
authors.initials: TT
authors.lastname: Cao
authors.forename: Anne
authors.initials: A
authors.lastname: Brelot
authors.forename: Mark
authors.initials: M
authors.lastname: von Zastrow
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D007668:Kidney; D046911:Macromolecular Substances; D051379:Mice; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000502:physiology*; D017465:Receptors, Opioid, delta / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D046911:Macromolecular Substances; D018343:Receptors, Adrenergic, beta-2; D017465:Receptors, Opioid, delta; D011994:Recombinant Proteins