Abstract
Cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-regulated chloride channel localized at apical cell membranes and exists in macromolecular complexes with a variety of signaling and transporter molecules. Here, we report that the multidrug resistance protein 4 (MRP4), a cAMP transporter, functionally and physically associates with CFTR. Adenosine-stimulated CFTR-mediated chloride currents are potentiated by MRP4 inhibition, and this potentiation is directly coupled to attenuated cAMP efflux through the apical cAMP transporter. CFTR single-channel recordings and FRET-based intracellular cAMP dynamics suggest that a compartmentalized coupling of cAMP transporter and CFTR occurs via the PDZ scaffolding protein, PDZK1, forming a macromolecular complex at apical surfaces of gut epithelia. Disrupting this complex abrogates the functional coupling of cAMP transporter activity to CFTR function. Mrp4 knockout mice are more prone to CFTR-mediated secretory diarrhea. Our findings have important implications for disorders such as inflammatory bowel disease and secretory diarrhea.
MeSH Terms
Animals
Cell Compartmentation/physiology
Cell Membrane/drug effects,metabolism
Cells, Cultured
Chloride Channels/metabolism
Cyclic AMP/analogs & derivatives,metabolism
Cystic Fibrosis Transmembrane Conductance Regulator/metabolism,physiology
Diarrhea/chemically induced
HT29 Cells
Humans
Intestinal Mucosa/metabolism
Mice
Mice, Knockout
Models, Biological
Multidrug Resistance-Associated Proteins/antagonists & inhibitors,genetics,metabolism
Propionates/pharmacology
Protein Binding
Quinolines/pharmacology
Time Factors
Chemicals
ABCC4 protein, human
CFTR protein, human
Chloride Channels
Multidrug Resistance-Associated Proteins
Propionates
Quinolines
Cystic Fibrosis Transmembrane Conductance Regulator
verlukast
Cyclic AMP
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Li Chunying
Department of Physiology, The University of Tennessee Health Science Center, 894 Union Avenue, 420 Nash, Memphis, TN 38163, USA.
Krishnamurthy Partha C
Penmatsa Himabindu
Marrs Kevin L
Wang Xue Qing
Zaccolo Manuela
Jalink Kees
Li Min
Nelson Deborah J
Schuetz John D
Naren Anjaparavanda P
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