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Domain-interface dynamics of CFTR revealed by stabilizing nanobodies

DOI: 10.1038/s41467-019-10714-y DOI Help

Authors: Maud Sigoillot (SFMB, Université Libre de Bruxelles (ULB)) , Marie Overtus (SFMB, Université Libre de Bruxelles (ULB)) , Magdalena Grodecka (SFMB, Université Libre de Bruxelles (ULB)) , Daniel Scholl (SFMB, Université Libre de Bruxelles (ULB)) , Abel Garcia-pino (Université Libre de Bruxelles (ULB)) , Toon Laeremans (Vrije Universiteit Brussel (VUB); VIB-VUB center for Structural Biology, VIB) , Lihua He (University of North Carolina-Chapel Hill) , Els Pardon (Vrije Universiteit Brussel (VUB); VIB-VUB center for Structural Biology, VIB) , Ellen Hildebrandt (Texas Tech University Health Sciences Center) , Ina Urbatsch (Texas Tech University Health Sciences Center) , Jan Steyaert (Vrije Universiteit Brussel (VUB); VIB-VUB center for Structural Biology, VIB) , John R. Riordan (University of North Carolina-Chapel Hill) , Cedric Govaerts (SFMB, Université Libre de Bruxelles (ULB))
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 10 , PAGES 2636

State: Published (Approved)
Published: June 2019
Diamond Proposal Number(s): 12718 , 12718

Open Access Open Access

Abstract: The leading cause of cystic fibrosis (CF) is the deletion of phenylalanine 508 (F508del) in the first nucleotide-binding domain (NBD1) of the cystic fibrosis transmembrane conductance regulator (CFTR). The mutation affects the thermodynamic stability of the domain and the integrity of the interface between NBD1 and the transmembrane domain leading to its clearance by the quality control system. Here, we develop nanobodies targeting NBD1 of human CFTR and demonstrate their ability to stabilize both isolated NBD1 and full-length protein. Crystal structures of NBD1-nanobody complexes provide an atomic description of the epitopes and reveal the molecular basis for stabilization. Furthermore, our data uncover a conformation of CFTR, involving detachment of NBD1 from the transmembrane domain, which contrast with the compact assembly observed in cryo-EM structures. This unexpected interface rearrangement is likely to have major relevance for CF pathogenesis but also for the normal function of CFTR and other ABC proteins.

Journal Keywords: Chloride channels; Membrane proteins; X-ray crystallograph

Subject Areas: Biology and Bio-materials, Medicine

Instruments: I02-Macromolecular Crystallography , I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography