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The β2-adrenergic receptor interacts with the Na+/H+-exchanger regulatory factor to control Na+/H+ exchange

Nature volume 392, pages 626–630 (1998)Cite this article

Abstract

Stimulation of β2-adrenergic receptors on the cell surface by adrenaline or noradrenaline leads to alterations in the metabolism, excitability, differentiation and growth of many cell types. These effects have traditionally been thought to be mediated exclusively by receptor activation of intracellular G proteins1. However, certain physiological effects of β2-adrenergic receptor stimulation, notably the regulation of cellular pH by modulation of Na+/H+ exchanger (NHE) function, do not seem to be entirely dependent on G-protein activation2,3,4,5,6,7. We report here a direct agonist-promoted association of the β2-adrenergic receptor with the Na+/H+ exchanger regulatory factor (NHERF), a protein that regulates the activity of the Na+/H+ exchanger type 3 (NHE3)8. NHERF binds to the β2-adrenergic receptor by means of a PDZ-domain-mediated interaction with the last few residues of the carboxy-terminal cytoplasmic domain of the receptor. Mutation ofthe final residue of the β2-adrenergic receptor from leucine toalanine abolishes the receptor's interaction with NHERF andalso markedly alters β2-adrenergic receptor regulation of NHE3 in cells without altering receptor-mediated activation of adenylyl cyclase. Our findings indicate that agonist-dependent β2-adrenergic receptor binding of NHERF plays a role in β2-adrenergic receptor-mediated regulation of Na+/H+ exchange.

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Figure 1: a, Purification of a β2-receptor tail interacting protein.
Figure 2: a, NHERF binds to the C-terminal 10 amino acids of the β2-receptor tail.
Figure 3: NHERF co-localizes in cells with full-length wild-type β2 receptor, but not with full-length L413A mutant β2 receptor, after receptor stimulation with agonist.
Figure 4: β2-receptor tail–GST fusion protein blocks the ability of NHERF to regulate NHE3.
Figure 5: Agonist stimulation of L413A mutant β2 receptors but not wild-type β2 receptors inhibits NHE3 in whole cells.

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Acknowledgements

We thank J. Shannon for peptide sequencing; T. Kurose for the β2 receptor tail GST fusion protein construct; N. Freedman for the Flag-tagged wild-type β2 receptor construct; J. Raymond for advice; G. Irons, D.Steplock and K. Tate for technical assistance; and D. Addison and M. Holben for help in preparing the manuscript. This work was supported in part by grants from the NIH to R.J.L. and E.J.W. and from theDuke Comprehensive Cancer Center to S.S.; C.W.C. is the recipient of a clinician–scientist award fromthe Department of Medicine at the University of Toronto; A.C. is a recipient of a postdoctoral fellowship from the Heart and Stroke Foundation of Canada; and S.G. is an international scholar of the Howard Hughes Medical Institute.

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Author notes

  1. Robert H. Stoffel

    Present address: Bristol Myers Squibb, Pharmaceutical Research Institute, Wallingford, Connecticut, 06492, USA

  2. Edward J. Weinman

    Present address: GI Division, Department of Medicine, Johns Hopkins University, Baltimore, Maryland, 21205, USA

Authors and Affiliations

  1. Departments of Medicine, Howard Hughes Medical Institute, Biochemistry and Cell Biology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Randy A. Hall, Richard T. Premont, Jeremy T. Blitzer, Julie A. Pitcher, Audrey Claing, Robert H. Stoffel, Larry S. Barak & Robert J. Lefkowitz

  2. Division of Cell Biology, Research Institute, Hospital for Sick Children, Toronto, M5G 1X8, Canada

    Chung-Wai Chow & Sergio Grinstein

  3. Department of Pharmacology and Cancer Biology, Duke University, Durham, 27710, North Carolina, USA

    Shirish Shenolikar

  4. Department of Medicine, Robert C. Byrd Health Science Center, West Virginia University, Morgantown, 26506, West Virginia

    Edward J. Weinman

  5. Department of Veteran Affairs Medical Center, Medical Service, Clarksburg, 26301, West Virginia, USA

    Edward J. Weinman

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Correspondence to Robert J. Lefkowitz.

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Hall, R., Premont, R., Chow, CW. et al. The β2-adrenergic receptor interacts with the Na+/H+-exchanger regulatory factor to control Na+/H+ exchange. Nature 392, 626–630 (1998). https://doi.org/10.1038/33458

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