Details

Autor(en) / Beteiligte

• Faure, Grazyna
• Bakouh, Naziha
• Lourdel, Stéphane
• Odolczyk, Norbert
• Premchandar, Aiswarya
• Servel, Nathalie
• Hatton, Aurélie
• Ostrowski, Maciej K.
• Xu, Haijin
• Saul, Frederick A.
• Moquereau, Christelle
• Bitam, Sara
• Pranke, Iwona
• Planelles, Gabrielle
• Teulon, Jacques
• Herrmann, Harald
• Roldan, Ariel
• Zielenkiewicz, Piotr
• Dadlez, Michal
• Lukacs, Gergely L.
• Sermet-Gaudelus, Isabelle
• Ollero, Mario
• Corringer, Pierre-Jean
• Edelman, Aleksander

Titel

Rattlesnake Phospholipase A2 Increases CFTR-Chloride Channel Current and Corrects ∆F508CFTR Dysfunction: Impact in Cystic Fibrosis

Ist Teil von

• Journal of molecular biology, 2016-07, Vol.428 (14), p.2898-2915

Ort / Verlag

England: Elsevier Ltd

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •The CB subunit of crotoxin is a novel binding partner for CFTR.•CB binding to CFTR potentiates chloride channel current in normal and ∆F508CFTR.•CB interaction with ∆F508CFTR corrects its cellular localization defect.•A structural model of the CB/ΔF508CFTR binding interface is presented.•CB–CFTR interaction constitutes a novel target of therapeutic strategies in CF.