Rattlesnake phospholipase A2 increases CFTR-chloride channel current and corrects ?F508CFTR dysfunction: impact in cystic fibrosis.
PBN-AR
Instytucja
Instytut Biochemii i Biofizyki Polskiej Akademii Nauk
Informacje podstawowe
Główny język publikacji
EN
Czasopismo
Journal of Molecular Biology
ISSN
0022-2836
EISSN
1089-8638
Wydawca
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI
Rok publikacji
2016
Numer zeszytu
14
Strony od-do
2898-2915
Numer tomu
428
Identyfikator DOI
Liczba arkuszy
Autorzy
Pozostali autorzy
+ 20
Słowa kluczowe
EN
chloride channel;
crotoxin;
cystic fibrosis;
phospholipase A(2);
biomolecular interactions
Streszczenia
Język
EN
Treść
Deletion of Phe508 in the nucleotide binding domain (∆ F508–NBD1) of the cystic fibrosis transmembrane regulator (CFTR; a cyclic AMP-regulated chloride channel) is the most frequent mutation associated with cystic fibrosis. This mutation affects the maturation and gating of CFTR protein. The search for new high-affinity ligands of CFTR acting as dual modulators (correctors/activators) presents a major challenge in the pharmacology of cystic fibrosis. Snake venoms are a rich source of natural multifunctional proteins, potential binders of ion channels. In this study, we identified the CB subunit of crotoxin from Crotalus durissus terrificus as a new ligand and allosteric modulator of CFTR. We showed that CB interacts with NBD1 of both wild type and ∆ F508CFTR and increases their chloride channel currents. The potentiating effect of CB on CFTR activity was demonstrated using electrophysiological techniques in Xenopus laevis oocytes, in CFTR–HeLa cells, and ex vivo in mouse colon tissue. The correcting effect of CB was shown by functional rescue of CFTR activity after 24-h ΔF508CFTR treatments with CB. Moreover, the presence of fully glycosylated CFTR was observed. Molecular docking allowed us to propose a model of the complex involving of the ABCβ and F1-like ATP-binding subdomains of ΔF508–NBD1. Hydrogen–deuterium exchange analysis confirmed stabilization in these regions, also showing allosteric stabilization in two other distal regions. Surface plasmon resonance competition studies showed that CB disrupts the ∆ F508CFTR–cytokeratin 8 complex, allowing for the escape of ∆ F508CFTR from degradation. Therefore CB, as a dual modulator of ΔF508CFTR, constitutes a template for the development of new anti-CF agents.
Inne
System-identifier
PX-57d025c6c2dcd20f1cf3e9f9
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