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Nat Commun. 2019 Jun 14;10(1):2636. doi: 10.1038/s41467-019-10714-y.

Domain-interface dynamics of CFTR revealed by stabilizing nanobodies.

Author information

1
SFMB, Université Libre de Bruxelles (ULB), CP206/02, Boulevard du Triomphe, building BC, B-1050, Brussels, Belgium.
2
Laboratoire de Microbiologie Moléculaire et Cellulaire, ULB CP300, rue des Professeurs Jeener et Brachet 12, B-6041, Charleroi, Belgium.
3
Structural Biology Brussels, Vrije Universiteit Brussel (VUB), Pleinlaan 2, B-1050, Brussels, Belgium.
4
VIB-VUB center for Structural Biology, VIB, Pleinlaan 2, B-1050, Brussels, Belgium.
5
Department of Biochemistry and Biophysics and Cystic Fibrosis Center, University of North Carolina-Chapel Hill, Chapel Hill, NC, 27599, USA.
6
Department of Cell Biology and Biochemistry and Center for Membrane Protein Research, Texas Tech University Health Sciences Center, 3601 4th Street, Stop 6540, Lubbock, TX, 79430, USA.
7
SFMB, Université Libre de Bruxelles (ULB), CP206/02, Boulevard du Triomphe, building BC, B-1050, Brussels, Belgium. Cedric.Govaerts@ulb.ac.be.

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.

PMID:
31201318
PMCID:
PMC6572788
DOI:
10.1038/s41467-019-10714-y
[Indexed for MEDLINE]
Free PMC Article

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