Nanomechanics combined with HDX reveals allosteric drug binding sites of CFTR NBD1

Cystic fibrosis (CF) is a frequent genetic disease in Caucasians that is caused by the deletion of F508 (Delta F508) in the nucleotide binding domain 1 (NBD1) of the CF transmembrane conductance regulator (CFTR). The Delta F508 compromises the folding energetics of the NBD1, as well as the folding of three other CFTR domains. Combination of FDA approved corrector molecules can efficiently but incompletely rescue the Delta F508-CFTR folding and stability defect. Thus, new pharmacophores that would reinstate the wildtype-like conformational stability of the Delta F508-NBD1 would be highly beneficial. The most prominent molecule, 5-bromoindole-3-acetic acid (BIA) that can thermally stabilize the NBD1 has low potency and efficacy. To gain insights into the NBD1 (un)folding dynamics and BIA binding site localization, we combined molecular dynamics (MD) simulations, atomic force spectroscopy (AFM) and hydrogen-deuterium exchange (HDX) experiments. We found that the NBD1 alpha-subdomain with three adjacent strands from the beta-subdomain plays an important role in early folding steps, when crucial non-native interactions are formed via residue F508. Our AFM and HDX experiments showed that BIA associates with this alpha-core region and increases the resistance of the Delta F508-NBD1 against mechanical unfolding, a phenomenon that could be exploited in future developments of folding correctors. (C) 2022 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Automated summary

Cystic fibrosis is a common genetic disease in Caucasians caused by the deletion of F508 in the CFTR gene. Current treatments can partially correct the folding and stability defect, but new pharmacophores are needed to restore the wildtype-like conformational stability of the mutated protein. The molecule BIA can thermally stabilize the NBD1 region and increase its resistance to mechanical unfolding, suggesting its potential as a folding corrector.