Characterization of the molecular interaction between caveolin-1 and the P2X receptors 4 and 7 in E10 mouse lung alveolar epithelial cells

The International Journal of Biochemistry & Cell Biology
K BarthM Kasper

Abstract

P2X(4) and P2X(7) receptors are abundantly expressed in alveolar epithelial cells, and are thought to play a role in regulating fluid haemostasis. Here, we analyzed the expression and localization of the P2X(4)R, and characterized the interaction between Cav-1 and both P2X(4)R and P2X(7)R in the mouse alveolar epithelial cell line E10. Using the biotinylation assay, we found that only glycosylated P2X(4)R is exposed at the cell surface. Triton X-100 solubility experiments and sucrose gradient centrifugation revealed that P2X(4)R was partially localized in Cav-1 rich membrane fractions. Cholesterol depletion with Mbeta-CD displaced Cav-1 and P2X(4)R from the low-density to the high-density fractions. Suppression of Cav-1 protein expression using short hairpin RNAs resulted in a large reduction in P2X(4)R levels. Double immunofluorescence showed that P2X(4)R and Cav-1 partially colocalize in vitro. Using the GST pull-down assay, we showed that Cav-1 interacts in vitro with both P2X(4)R and P2X(7)R. Co-immunoprecipitation experiments confirmed the interaction between P2X(7)R and Cav-1. ATP stimulation increased the level of P2X(4)R in the lipid raft/caveolae fraction, whereas Cav-1 content remained constant. Our results support re...Continue Reading

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Citations

Jun 22, 2010·Histochemistry and Cell Biology·K BarthM Kasper
Feb 1, 2011·Histochemistry and Cell Biology·Stefan Hübner, Athina Efthymiadis
May 16, 2012·Histochemistry and Cell Biology·Claudia PflegerKathrin Barth
Jan 10, 2012·Purinergic Signalling·Jing Li, Samuel J Fountain
Oct 4, 2008·Annual Review of Physiology·Douglas C EatonLucky Jain
Jun 5, 2010·American Journal of Respiratory and Critical Care Medicine·Nicolas RiteauIsabelle Couillin
Apr 24, 2014·Journal of Molecular Cell Biology·Xu-Qiao ChenLan Bao
Apr 21, 2010·Cellular and Molecular Life Sciences : CMLS·Karina WeinholdKathrin Barth
Oct 13, 2010·The International Journal of Biochemistry & Cell Biology·Annett LingeKathrin Barth
Oct 5, 2014·The Journal of Biological Chemistry·Lucy E RobinsonRuth D Murrell-Lagnado
Aug 14, 2012·Pharmacological Reviews·Geoffrey BurnstockJean-Pierre Timmermans
Oct 14, 2017·Bioscience Reports·Michael Kasper, Kathrin Barth
Oct 2, 2020·Arteriosclerosis, Thrombosis, and Vascular Biology·Brandon G ShokoplesErnesto L Schiffrin
Aug 24, 2019·Frontiers in Molecular Neuroscience·Robin KoppAnnette Nicke
Jun 5, 2013·Médecine sciences : M/S·Stéphanie PochetJean-Paul Dehaye

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