WS4.4 Restoration of F508Δ-CFTR trafficking and function by liposome-mediated delivery of antibodies against cytokeratin 8

Chatin, Benoît; Mevel, M.; Edelman, A.; Pitard, B.

doi:10.1016/s1569-1993(13)60022-3

Cited by 1 publication

(2 citation statements)

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“…First, the majority of proteins that interact with IFs target the head domain (desmoplakin, plectin, fimbrin, and S100α/β). Second, antibodies directed against the N‐terminal fragment of K8 (but not against its C‐terminus) introduced into ΔF508‐CFTR‐expressing HeLa cells reportedly lead to the functional expression of mutated CFTR . Third, the dynamic properties of K8 displayed several fragments with high flexibility, with the head region among the regions of highest flexibility .…”

Section: Resultsmentioning

confidence: 99%

“…Second, antibodies directed against the N-terminal fragment of K8 (but not against its C-terminus) introduced into DF508-CFTR-expressing HeLa cells reportedly lead to the functional expression of mutated CFTR. 29 Third, the dynamic properties of K8 displayed several fragments with high flexibility, with the head region among the regions of highest flexibility. 15 Based on these observations, three peptides from the head region and a short N-terminal fragment of coil 1A of K8 13 were selected for further interaction studies and synthesized: In the DF508-NBD1-Peptide 1 complex, we observed additional stabilization in the regions 400-408 from RI, and in three other peptides between positions 550-650 of the F1-like core subdomain.…”

Section: Identification Of K8 Fragments That Interact With Nbd1mentioning

confidence: 96%

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New insights into interactions between the nucleotide‐binding domain of CFTR and keratin 8

et al. 2017

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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 (ΔF508), and this interaction hampers the biogenesis of functional ΔF508-CFTR and its insertion into the plasma membrane. Interruption of this interaction may constitute a new therapeutic target for CF patients bearing the ΔF508 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 ΔF508 mutation, we used hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS) on recombinant wild-type (wt) NBD1 and ΔF508-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 ΔF508-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 ΔF508-NBD1, which is supported by an additional binding site located in the vicinity of the ΔF508 mutation in NBD1.

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