Role of interaction and nucleoside diphosphate kinase B in regulation of the cystic fibrosis transmembrane conductance regulator function by cAMP-dependent protein kinase A

Lee A. Borthwick, Mathieu Kerbiriou, Christopher J. Taylor, Giorgio Cozza, Ioan Lascu, Edith H. Postel, Diane Cassidy, Pascal Trouvé, Anil Mehta, Louise Robson, Richmond Muimo (Lead / Corresponding author)

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    Abstract

    Cystic fibrosis results from mutations in the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-dependent protein kinase A (PKA) and ATP-regulated chloride channel. Here, we demonstrate that nucleoside diphosphate kinase B (NDPK-B, NM23-H2) forms a functional complex with CFTR. In airway epithelia forskolin/IBMX significantly increases NDPK-B co-localisation with CFTR whereas PKA inhibitors attenuate complex formation. Furthermore, an NDPK-B derived peptide (but not its NDPK-A equivalent) disrupts the NDPK-B/CFTR complex in vitro (19-mers comprising amino acids 36-54 from NDPK-B or NDPK-A). Overlay (Far-Western) and Surface Plasmon Resonance (SPR) analysis both demonstrate that NDPK-B binds CFTR within its first nucleotide binding domain (NBD1, CFTR amino acids 351-727). Analysis of chloride currents reflective of CFTR or outwardly rectifying chloride channels (ORCC, DIDS-sensitive) showed that the 19-mer NDPK-B peptide (but not its NDPK-A equivalent) reduced both chloride conductances. Additionally, the NDPK-B (but not NDPK-A) peptide also attenuated acetylcholine-induced intestinal short circuit currents. In silico analysis of the NBD1/NDPK-B complex reveals an extended interaction surface between the two proteins. This binding zone is also target of the 19-mer NDPK-B peptide, thus confirming its capability to disrupt NDPK-B/CFTR complex. We propose that NDPK-B forms part of the complex that controls chloride currents in epithelia.

    Original languageEnglish
    Article numbere0149097
    Number of pages25
    JournalPLoS ONE
    Volume11
    Issue number3
    DOIs
    Publication statusPublished - 7 Mar 2016

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  • Cite this

    Borthwick, L. A., Kerbiriou, M., Taylor, C. J., Cozza, G., Lascu, I., Postel, E. H., Cassidy, D., Trouvé, P., Mehta, A., Robson, L., & Muimo, R. (2016). Role of interaction and nucleoside diphosphate kinase B in regulation of the cystic fibrosis transmembrane conductance regulator function by cAMP-dependent protein kinase A. PLoS ONE, 11(3), [e0149097]. https://doi.org/10.1371/journal.pone.0149097