New insights into interactions between the nucleotide-binding domain of CFTR and keratin 8.
PBN-AR
Instytucja
Instytut Biochemii i Biofizyki Polskiej Akademii Nauk
Informacje podstawowe
Główny język publikacji
EN
Czasopismo
Protein science : a publication of the Protein Society
ISSN
EISSN
1469-896X
Wydawca
WILEY-BLACKWEL
DOI
Rok publikacji
2016
Numer zeszytu
2
Strony od-do
343-354
Numer tomu
26
Identyfikator DOI
Liczba arkuszy
Autorzy
Pozostali autorzy
+ 9
Słowa kluczowe
EN
CFTR;
NBD1;
cystic fibrosis;
hydrogen-deuterium exchange mass spectrometry;
keratin 8;
protein structure
Streszczenia
Język
EN
Treść
The intermediate filament protein keratin 8 (K8) interacts with the nucleotide-binding domain 1 (NBD1) of the cystic fibrosis transmembrane regulator (CFTR) with phenylalanine 508 deletion (DeltaF508), and this interaction hampers the biogenesis of functional DeltaF508-CFTR and its insertion into the plasma membrane. Interruption of this interaction may constitute a new therapeutic target for cystic fibrosis patients bearing the DeltaF508 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 DeltaF508 mutation, we used hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS) on recombinant wild-type (wt) NBD1 and DeltaF508-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 DeltaF508-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 DeltaF508-NBD1, which is supported by an additional binding site located in the vicinity of the DeltaF508 mutation in NBD1.
Inne
System-identifier
PX-5893472282ce4a248cecdc54
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