Journal Article

Domain-interface dynamics of CFTR revealed by stabilizing nanobodies

Grodecka, Magdalena, Govaerts, Cedric, He, Lihua, Riordan, John R., Pardon, Els, Urbatsch, Ina, Steyaert, Jan, Garcia-Pino, Abel, Sigoillot, Maud, Scholl, Daniel, Overtus, Marie, Hildebrandt, Ellen, Laeremans, Toon

2019

Overview

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. The leading cause of cystic fibrosis is the deletion of phenylalanine 508 (F508del) in the first nucleotide-binding domain (NBD1) of the cystic fibrosis transmembrane conductance regulator (CFTR). Here authors we develop nanobodies targeting NBD1 of human CFTR and demonstrate their ability to stabilize both isolated NBD1 and full-length protein.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 101/1 / 631/45/269/1147 / 631/45/535/1266 / 631/45/612/1237 / 82/80 / 82/83 / 9/10 / Conductance / Control systems / Crystal structure / Crystallography, X-Ray / Cystic fibrosis / Cystic Fibrosis - genetics / Cystic Fibrosis Transmembrane Conductance Regulator - genetics / Cystic Fibrosis Transmembrane Conductance Regulator - isolation & purification / Cystic Fibrosis Transmembrane Conductance Regulator - metabolism / Epitopes / Gene deletion / Humanities and Social Sciences / Humans / Interface stability / Models, Molecular / multidisciplinary / Mutation / Nanobodies / Nucleotides / Pathogenesis / Phenylalanine / Protein Folding / Protein Interaction Domains and Motifs - genetics / Protein Stability / Protein structure / Protein Structure, Tertiary - genetics / Proteins / Quality control / Recombinant Proteins - genetics / Recombinant Proteins - isolation & purification / Recombinant Proteins - metabolism / Science / Science (multidisciplinary) / Sequence Deletion / Single-Domain Antibodies - metabolism