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Small-angle X-ray scattering study of the ATP modulation of the structural features of the nucleotide binding domains of the CFTR in solution

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Abstract

Nucleotide binding domains (NBD1 and NBD2) of the cystic fibrosis transmembrane conductance (CFTR), the defective protein in cystic fibrosis, are responsible for controlling the gating of the chloride channel and are the putative binding site for several candidate drugs in the disease treatment. We studied the structural properties of recombinant NBD1, NBD2, and an equimolar NBD1/NBD2 mixture in solution by small-angle X-ray scattering. We demonstrated that NBD1 or NBD2 alone have an overall structure similar to that observed for crystals. Application of 2 mM ATP induces a dimerization of NBD1 but does not modify the NBD2 monomeric conformation. An equimolar mixture of NBD1/NBD2 in solution shows a dimeric conformation, and the application of ATP to the solution causes a conformational change in the NBD1/NBD2 complex into a tight heterodimer. We hypothesize that a similar conformation change occurs in situ and that transition is part of the gating mechanism. To our knowledge, this is the first direct observation of a conformational change of the NBD1/NBD2 interaction by ATP. This information may be useful to understand the physiopathology of cystic fibrosis.

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Acknowledgments

We thank the ESRF for provision of synchrotron radiation facilities, and we would like to thank Petra Pernod for assistance in using beamline ID14-EH3. We thank Olga Zegarra, Gino Galietta, Paola Vergani, and Ilaria Zanardi for their comments. This project was supported by the Fondazione Ricerca Fibrosi Cistica (grant #2/2008), Mille bambini a Via Margutta–onlus, Blunotte and Lega Italiana FC–Associazione Toscana Onlus.

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  1. Istituto di Biofisica, Consiglio Nazionale delle Ricerche, Via De Marini, 6, 16149, Genoa, Italy

    Lauretta Galeno, Elena Galfrè & Oscar Moran

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  1. Lauretta Galeno
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  2. Elena Galfrè
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  3. Oscar Moran
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Correspondence to Oscar Moran.

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Galeno, L., Galfrè, E. & Moran, O. Small-angle X-ray scattering study of the ATP modulation of the structural features of the nucleotide binding domains of the CFTR in solution. Eur Biophys J 40, 811–824 (2011). https://doi.org/10.1007/s00249-011-0692-5

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  • DOI: https://doi.org/10.1007/s00249-011-0692-5

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