PT  - JOURNAL ARTICLE
AU  - Onofrio Laselva
AU  - Steven Molinski
AU  - Valeria Casavola
AU  - Christine E. Bear
TI  - Correctors of the Major Cystic Fibrosis Mutant Interact through Membrane-Spanning Domains
AID  - 10.1124/mol.118.111799
DP  - 2018 Jun 01
TA  - Molecular Pharmacology
PG  - 612--618
VI  - 93
IP  - 6
4099  - http://molpharm.aspetjournals.org/content/93/6/612.short
4100  - http://molpharm.aspetjournals.org/content/93/6/612.full
SO  - Mol Pharmacol2018 Jun 01; 93
AB  - The most common cystic fibrosis causing mutation is deletion of phenylalanine at position 508 (F508del), a mutation that leads to protein misassembly with defective processing. Small molecule corrector compounds: VX-809 or Corr-4a (C4) partially restores processing of the major mutant. These two prototypical corrector compounds cause an additive effect on F508del/cystic fibrosis transmembrane conductance regulator (CFTR) processing, and hence were proposed to act through distinct mechanisms: VX-809 stabilizing the first membrane-spanning domain (MSD) 1, and C4 acting on the second half of the molecule [consisting of MSD2 and/or nucleotide binding domain (NBD) 2]. We confirmed the effect of VX-809 in enhancing the stability of MSD1 and showed that it also allosterically modulates MSD2 when coexpressed with MSD1. We showed for the first time that C4 stabilizes the second half of the CFTR protein through its action on MSD2. Given the allosteric effect of VX-809 on MSD2, we were prompted to test the hypothesis that the two correctors interact in the full-length mutant protein. We did see evidence supporting their interaction in the full-length F508del-CFTR protein bearing secondary mutations targeting domain:domain interfaces. Disruption of the MSD1:F508del-NBD1 interaction (R170G) prevented correction by both compounds, pointing to the importance of this interface in processing. On the other hand, stabilization of the MSD2:F508del-NBD1 interface (by introducing R1070W) led to a synergistic effect of the compound combination on the total abundance of both the immature and mature forms of the protein. Together, these findings suggest that the two correctors interact in stabilizing the complex of MSDs in F508del-CFTR.