A Macromolecular Complex of Beta 2 Adrenergic Receptor, CFTR, and Ezrin/radixin/moesin-binding Phosphoprotein 50 is Regulated by PKA

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2002 Dec 28

PMID 12502786

Citations 93

Authors

Anjaparavanda P Naren

Bryan Cobb

Chunying Li

Koushik Roy

David Nelson

Ghanshyam D Heda

Jie Liao

Kevin L Kirk

Eric J Sorscher

John Hanrahan

John P Clancy

Affiliations

Soon will be listed here.

Abstract

It has been demonstrated previously that both the cystic fibrosis transmembrane conductance regulator (CFTR) and beta(2) adrenergic receptor (beta(2)AR) can bind ezrinradixinmoesin-binding phosphoprotein 50 (EBP50, also referred to as NHERF) through their PDZ motifs. Here, we show that beta(2) is the major adrenergic receptor isoform expressed in airway epithelia and that it colocalizes with CFTR at the apical membrane. beta(2)AR stimulation increases CFTR activity, in airway epithelial cells, that is glybenclamide sensitive. Deletion of the PDZ motif from CFTR uncouples the channel from the receptor both physically and functionally. This uncoupling is specific to the beta(2)AR receptor and does not affect CFTR coupling to other receptors (e.g., adenosine receptor pathway). Biochemical studies demonstrate the existence of a macromolecular complex involving CFTR-EBP50-beta(2)AR through PDZ-based interactions. Assembly of the complex is regulated by PKA-dependent phosphorylation. Deleting the regulatory domain of CFTR abolishes PKA regulation of complex assembly. This report summarizes a macromolecular signaling complex involving CFTR, the implications of which may be relevant to CFTR-dysfunction diseases.

Citing Articles

Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators.

Botelho H, Lopes-Pacheco M, Pinto M, Railean V, Pankonien I, Caleiro M iScience. 2025; 28(3):111942.

PMID: 40040803 PMC: 11876911. DOI: 10.1016/j.isci.2025.111942.

Update on the Role of β2AR and TRPV1 in Respiratory Diseases.

Manti S, Gambadauro A, Galletta F, Ruggeri P, Piedimonte G Int J Mol Sci. 2024; 25(19).

PMID: 39408565 PMC: 11477158. DOI: 10.3390/ijms251910234.

Inflammation in the COVID-19 airway is due to inhibition of CFTR signaling by the SARS-CoV-2 spike protein.

Caohuy H, Eidelman O, Chen T, Mungunsukh O, Yang Q, Walton N Sci Rep. 2024; 14(1):16895.

PMID: 39043712 PMC: 11266487. DOI: 10.1038/s41598-024-66473-4.

Functional Consequences of CFTR Interactions in Cystic Fibrosis.

Ramananda Y, Naren A, Arora K Int J Mol Sci. 2024; 25(6).

PMID: 38542363 PMC: 10970640. DOI: 10.3390/ijms25063384.

It Takes Two to Tango! Protein-Protein Interactions behind cAMP-Mediated CFTR Regulation.

Murabito A, Bhatt J, Ghigo A Int J Mol Sci. 2023; 24(13).

PMID: 37445715 PMC: 10341646. DOI: 10.3390/ijms241310538.

References

Smith J, Karp P, Welsh M . Defective fluid transport by cystic fibrosis airway epithelia. J Clin Invest. 1994; 93(3):1307-11. PMC: 294089. DOI: 10.1172/JCI117087. View

Huang P, Lazarowski E, Tarran R, Milgram S, Boucher R, Stutts M . Compartmentalized autocrine signaling to cystic fibrosis transmembrane conductance regulator at the apical membrane of airway epithelial cells. Proc Natl Acad Sci U S A. 2001; 98(24):14120-5. PMC: 61178. DOI: 10.1073/pnas.241318498. View

Winter M, Welsh M . Stimulation of CFTR activity by its phosphorylated R domain. Nature. 1997; 389(6648):294-6. DOI: 10.1038/38514. View

Barak L, Tiberi M, Freedman N, Kwatra M, Lefkowitz R, Caron M . A highly conserved tyrosine residue in G protein-coupled receptors is required for agonist-mediated beta 2-adrenergic receptor sequestration. J Biol Chem. 1994; 269(4):2790-5. View

Chang X, Tabcharani J, Hou Y, Jensen T, Kartner N, Alon N . Protein kinase A (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites. J Biol Chem. 1993; 268(15):11304-11. View

Clancy J, Ruiz F, Sorscher E . Adenosine and its nucleotides activate wild-type and R117H CFTR through an A2B receptor-coupled pathway. Am J Physiol. 1999; 276(2):C361-9. DOI: 10.1152/ajpcell.1999.276.2.C361. View

Hall R, Premont R, Chow C, Blitzer J, Pitcher J, Claing A . The beta2-adrenergic receptor interacts with the Na+/H+-exchanger regulatory factor to control Na+/H+ exchange. Nature. 1998; 392(6676):626-30. DOI: 10.1038/33458. View

Prince L, Tousson A, Marchase R . Cell surface labeling of CFTR in T84 cells. Am J Physiol. 1993; 264(2 Pt 1):C491-8. DOI: 10.1152/ajpcell.1993.264.2.C491. View

Fouassier L, Yun C, Fitz J, DOCTOR R . Evidence for ezrin-radixin-moesin-binding phosphoprotein 50 (EBP50) self-association through PDZ-PDZ interactions. J Biol Chem. 2000; 275(32):25039-45. DOI: 10.1074/jbc.C000092200. View

10.

Raghuram V, Mak D, Foskett J . Regulation of cystic fibrosis transmembrane conductance regulator single-channel gating by bivalent PDZ-domain-mediated interaction. Proc Natl Acad Sci U S A. 2001; 98(3):1300-5. PMC: 14749. DOI: 10.1073/pnas.98.3.1300. View

11.

Matsui H, Davis C, Tarran R, Boucher R . Osmotic water permeabilities of cultured, well-differentiated normal and cystic fibrosis airway epithelia. J Clin Invest. 2000; 105(10):1419-27. PMC: 315457. DOI: 10.1172/JCI4546. View

12.

Green S, Spasoff A, Coleman R, Johnson M, Liggett S . Sustained activation of a G protein-coupled receptor via "anchored" agonist binding. Molecular localization of the salmeterol exosite within the 2-adrenergic receptor. J Biol Chem. 1996; 271(39):24029-35. DOI: 10.1074/jbc.271.39.24029. View

13.

Hanrahan J, Tabcharani J, Becq F, Mathews C, Augustinas O, Jensen T . Function and dysfunction of the CFTR chloride channel. Soc Gen Physiol Ser. 1995; 50:125-37. View

14.

Wang S, Yue H, Derin R, Guggino W, Li M . Accessory protein facilitated CFTR-CFTR interaction, a molecular mechanism to potentiate the chloride channel activity. Cell. 2000; 103(1):169-79. DOI: 10.1016/s0092-8674(00)00096-9. View

15.

Zhu T, Dahan D, Evagelidis A, Zheng S, Luo J, Hanrahan J . Association of cystic fibrosis transmembrane conductance regulator and protein phosphatase 2C. J Biol Chem. 1999; 274(41):29102-7. DOI: 10.1074/jbc.274.41.29102. View

16.

Chan H, Fong S, So S, Chung Y, Wong P . Stimulation of anion secretion by beta-adrenoceptors in the mouse endometrial epithelium. J Physiol. 1997; 501 ( Pt 3):517-25. PMC: 1159453. DOI: 10.1111/j.1469-7793.1997.517bm.x. View

17.

Prince L, Peter K, Hatton S, Zaliauskiene L, Cotlin L, Clancy J . Efficient endocytosis of the cystic fibrosis transmembrane conductance regulator requires a tyrosine-based signal. J Biol Chem. 1999; 274(6):3602-9. DOI: 10.1074/jbc.274.6.3602. View

18.

Clancy B, Czech M . Hexose transport stimulation and membrane redistribution of glucose transporter isoforms in response to cholera toxin, dibutyryl cyclic AMP, and insulin in 3T3-L1 adipocytes. J Biol Chem. 1990; 265(21):12434-43. View

19.

Rousseau G, Nantel F, Bouvier M . Distinct receptor domains determine subtype-specific coupling and desensitization phenotypes for human beta1- and beta2-adrenergic receptors. Mol Pharmacol. 1996; 49(4):752-60. View

20.

Naren A, Di A, Cormet-Boyaka E, Boyaka P, McGhee J, Zhou W . Syntaxin 1A is expressed in airway epithelial cells, where it modulates CFTR Cl(-) currents. J Clin Invest. 2000; 105(3):377-86. PMC: 377449. DOI: 10.1172/JCI8631. View