New insights into interactions between the nucleotide-binding domain of CFTR and keratin 8
Premchandar A, Kupniewska A, Bonna A, Faure G, Fraczyk T, Roldan A, Hoffmann B, Da Cunha MF, Herrmann H, Lukacs GL, Edelman A, Dadlez M (2017)
Publication Type: Journal article
Publication year: 2017
Journal
Book Volume: 26
Pages Range: 343-354
Journal Issue: 2
DOI: 10.1002/pro.3086
Abstract
The intermediate filament protein keratin 8 (K8) interacts with the nucleotide-binding domain 1 (NBD1) of the cystic fibrosis (CF) transmembrane regulator (CFTR) with phenylalanine 508 deletion (?F508), and this interaction hampers the biogenesis of functional ?F508-CFTR and its insertion into the plasma membrane. Interruption of this interaction may constitute a new therapeutic target for CF patients bearing the ?F508 mutation. Here, we aimed to determine the binding surface between these two proteins, to facilitate the design of the interaction inhibitors. To identify the NBD1 fragments perturbed by the ?F508 mutation, we used hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS) on recombinant wild-type (wt) NBD1 and ?F508-NBD1 of CFTR. We then performed the same analysis in the presence of a peptide from the K8 head domain, and extended this investigation using bioinformatics procedures and surface plasmon resonance, which revealed regions affected by the peptide binding in both wt-NBD1 and ?F508-NBD1. Finally, we performed HDX-MS analysis of the NBD1 molecules and full-length K8, revealing hydrogen-bonding network changes accompanying complex formation. In conclusion, we have localized a region in the head segment of K8 that participates in its binding to NBD1. Our data also confirm the stronger binding of K8 to ?F508-NBD1, which is supported by an additional binding site located in the vicinity of the ?F508 mutation in NBD1.
Additional Organisation(s)
Involved external institutions
McGill University
Canada (CA)
Polska Akademia Nauk (PAN) / Polish Academy of Sciences
Poland (PL)
University of Paris 5 - René Descartes / Université Paris V René Descartes
France (FR)
Institut Pasteur
France (FR)
University of Paris 6 - Pierre et Marie Curie / Université Paris VI Pierre et Marie Curie (UPMC)
France (FR)
Canada (CA)
Polska Akademia Nauk (PAN) / Polish Academy of Sciences
Poland (PL)
University of Paris 5 - René Descartes / Université Paris V René Descartes
France (FR)
Institut Pasteur
France (FR)
University of Paris 6 - Pierre et Marie Curie / Université Paris VI Pierre et Marie Curie (UPMC)
France (FR)
How to cite
APA:
Premchandar, A., Kupniewska, A., Bonna, A., Faure, G., Fraczyk, T., Roldan, A.,... Dadlez, M. (2017). New insights into interactions between the nucleotide-binding domain of CFTR and keratin 8. Protein Science, 26(2), 343-354. https://doi.org/10.1002/pro.3086
MLA:
Premchandar, Aiswarya, et al. "New insights into interactions between the nucleotide-binding domain of CFTR and keratin 8." Protein Science 26.2 (2017): 343-354.
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