Stabilizing rescued surface-localized δf508 CFTR by potentiation of its interaction with Na(+)/H(+) exchanger regulatory factor 1

Biochemistry
Kavisha AroraAnjaparavanda P Naren

Abstract

Cystic fibrosis (CF) is a recessive genetic disease caused by mutations in CFTR, a plasma-membrane-localized anion channel. The most common mutation in CFTR, deletion of phenylalanine at residue 508 (ΔF508), causes misfolding of CFTR resulting in little or no protein at the plasma membrane. The CFTR corrector VX-809 shows promise for treating CF patients homozygous for ΔF508. Here, we demonstrate the significance of protein-protein interactions in enhancing the stability of the ΔF508 CFTR mutant channel protein at the plasma membrane. We determined that VX-809 prolongs the stability of ΔF508 CFTR at the plasma membrane. Using competition-based assays, we demonstrated that ΔF508 CFTR interacts poorly with Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) compared to wild-type CFTR, and VX-809 significantly increased this binding affinity. We conclude that stabilized CFTR-NHERF1 interaction is a determinant of the functional efficiency of rescued ΔF508 CFTR. Our results demonstrate the importance of macromolecular-complex formation in stabilizing rescued mutant CFTR at the plasma membrane and suggest this to be foundational for the development of a new generation of effective CFTR-corrector-based therapeutics.

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Citations

Sep 23, 2016·Frontiers in Pharmacology·Miquéias Lopes-Pacheco
Apr 1, 2017·American Journal of Physiology. Lung Cellular and Molecular Physiology·Carol A BertrandRaymond A Frizzell
Oct 30, 2016·American Journal of Physiology. Lung Cellular and Molecular Physiology·Neeraj SharmaGarry R Cutting
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Jun 5, 2016·American Journal of Physiology. Lung Cellular and Molecular Physiology·Kavisha AroraAnjaparavanda P Naren
Mar 2, 2017·Cold Spring Harbor Perspectives in Biology·Curtis T Okamoto
Jul 10, 2019·Scientific Reports·Yunjie HuangAnjaparavanda P Naren
Oct 6, 2017·JCI Insight·Kavisha AroraAnjaparavanda P Naren
Oct 16, 2018·Frontiers in Pharmacology·Ryosuke Fukuda, Tsukasa Okiyoneda
Jun 29, 2018·The Journal of Physiological Sciences : JPS·Kanako Wakabayashi-NakaoYoshiro Sohma
Jul 18, 2019·Nature Communications·Kyu Shik MunAnjaparavanda P Naren
Nov 17, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Kavisha AroraAnjaparavanda P Naren
Nov 26, 2020·American Journal of Physiology. Lung Cellular and Molecular Physiology·Chloé GrebertClarisse Vandebrouck
May 6, 2021·American Journal of Physiology. Gastrointestinal and Liver Physiology·Kavisha AroraAnjaparavanda P Naren
Jun 4, 2020·Molecular Therapy. Methods & Clinical Development·Steven ErwoodEvgueni A Ivakine
Jan 22, 2022·Journal of Cell Science·Asmahan Abu-ArishJohn W Hanrahan

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Methods Mentioned

BETA
scraping
proximity ligation assay
Assay
AlphaScreen
immunoprecipitation
ubiquitination

Software Mentioned

ImageJ
SPT
AlphaScreen
SlideBook

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