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A kinase-regulated PDZ-domain interaction controls endocytic sorting of the β2-adrenergic receptor

Nature volume 401, pages 286–290 (1999)Cite this article

Abstract

A fundamental question in cell biology is how membrane proteins are sorted in the endocytic pathway. The sorting of internalized β2-adrenergic receptors between recycling endosomes and lysosomes is responsible for opposite effects on signal transduction and is regulated by physiological stimuli1,2. Here we describe a mechanism that controls this sorting operation, which is mediated by a family of conserved protein-interaction modules called PDZ domains3. The phosphoprotein EBP50 (for ezrin–radixin–moesin(ERM)-binding phosphoprotein-50)4 binds to the cytoplasmic tail of the β2-adrenergic receptor through a PDZ domain and to the cortical actin cytoskeleton through an ERM-binding domain. Disrupting the interaction of EBP50 with either domain or depolymerization of the actin cytoskeleton itself causes missorting of endocytosed β2-adrenergic receptors but does not affect the recycling of transferrin receptors. A serine residue at position 411 in the tail of the β2-adrenergic receptor is a substrate for phosphorylation by GRK-5 (for G-protein-coupled-receptor kinase-5) (ref. 5) and is required for interaction with EBP50 and for proper recycling of the receptor. Our results identify a new role for PDZ-domain-mediated protein interactions and for the actin cytoskeleton in endocytic sorting, and suggest a mechanism by which GRK-mediated phosphorylation could regulate membrane trafficking of G-protein-coupled receptors after endocytosis.

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Figure 1: β2-AR mutants are degraded.
Figure 2: Missorting of mutant β2-ARs to lysosomes.
Figure 3: Role of EBP50 and actin in β2-AR sorting.
Figure 4: Regulation of β2-AR sorting by GRK-5.

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Acknowledgements

We thank P. Tsao for generating the β2-ARt cell line and for advice and assistance with flow cytometric experiments; U. Klein for the β2-AR tail GST fusion protein; J. Benovic for GRK-2 and -5 cDNAs; and R. Kelly, P. Walter, F. Brodsky, P. Peluso and J. Wilhelm for valuable discussion. T.T.C. is supported by an NIH Institutional Training Grant. These studies were supported by the NIH.

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, 94143, California, USA

    Tracy T. Cao & Heather W. Deacon

  2. Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York, 14853, USA

    David Reczek & Anthony Bretscher

  3. Program in Cell Biology, Cellular and Molecular Pharmacology and Psychiatry, University of California, San Francisco, San Francisco, California, 94143, USA

    Mark von Zastrow

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Correspondence to Mark von Zastrow.

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Cao, T., Deacon, H., Reczek, D. et al. A kinase-regulated PDZ-domain interaction controls endocytic sorting of the β2-adrenergic receptor. Nature 401, 286–290 (1999). https://doi.org/10.1038/45816

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