Corrector VX-809 promotes interactions between cytoplasmic loop one and the first nucleotide-binding domain of CFTR

TW Loo, DM Clarke - Biochemical Pharmacology, 2017 - Elsevier
TW Loo, DM Clarke
Biochemical Pharmacology, 2017Elsevier
A large number of correctors have been identified that can partially repair defects in folding,
stability and trafficking of CFTR processing mutants that cause cystic fibrosis (CF). The best
corrector, VX-809 (Lumacaftor), has shown some promise when used in combination with a
potentiator (Ivacaftor). Understanding the mechanism of VX-809 is essential for development
of better correctors. Here, we tested our prediction that VX-809 repairs folding and
processing defects of CFTR by promoting interactions between the first cytoplasmic loop …
Abstract
A large number of correctors have been identified that can partially repair defects in folding, stability and trafficking of CFTR processing mutants that cause cystic fibrosis (CF). The best corrector, VX-809 (Lumacaftor), has shown some promise when used in combination with a potentiator (Ivacaftor). Understanding the mechanism of VX-809 is essential for development of better correctors. Here, we tested our prediction that VX-809 repairs folding and processing defects of CFTR by promoting interactions between the first cytoplasmic loop (CL1) of transmembrane domain 1 (TMD1) and the first nucleotide-binding domain (NBD1). To investigate whether VX-809 promoted CL1/NBD1 interactions, we performed cysteine mutagenesis and disulfide cross-linking analysis of Cys-less TMD1 (residues 1–436) and ΔTMD1 (residues 437–1480; NBD1-R-TMD2-NBD2) truncation mutants. It was found that VX-809, but not bithiazole correctors, promoted maturation (exited endoplasmic reticulum for addition of complex carbohydrate in the Golgi) of the ΔTMD1 truncation mutant only when it was co-expressed in the presence of TMD1. Expression in the presence of VX-809 also promoted cross-linking between R170C (in CL1 of TMD1 protein) and L475C (in NBD1 of the ΔTMD1 truncation protein). Expression of the ΔTMD1 truncation mutant in the presence of TMD1 and VX-809 also increased the half-life of the mature protein in cells. The results suggest that the mechanism by which VX-809 promotes maturation and stability of CFTR is by promoting CL1/NBD1 interactions.
Elsevier