Abstract
The devastating inherited disease cystic fibrosis (CF) is caused by mutations of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel. The recent approval of the CFTR potentiator drug ivacaftor (Vx-770) for the treatment of CF patients has marked the advent of causative CF therapy. Currently, thousands of patients are being treated with the drug, and its molecular mechanism of action is under intensive investigation. Here we determine the solubility profile and true stimulatory potency of Vx-770 towards wild-type (WT) and mutant human CFTR channels in cell-free patches of membrane. We find that its aqueous solubility is ~200-fold lower (~60 nanomolar), whereas the potency of its stimulatory effect is >100-fold higher, than reported, and is unexpectedly fully reversible. Strong, but greatly delayed, channel activation by picomolar Vx-770 identifies multiple sequential slow steps in the activation pathway. These findings provide solid guidelines for the design of in vitro studies using Vx-770.
Article and author information
Author details
Funding
Cystic Fibrosis Foundation (CSANAD17G0)
- László Csanády
Magyar Tudományos Akadémia (Lendület grant LP2017-14/2017)
- László Csanády
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of Semmelweis University (last approved 06-30-2016, expiration 06-30-2021).
Reviewing Editor
- Leon D Islas, Universidad Nacional Autónoma de México, Mexico
Publication history
- Received: February 27, 2019
- Accepted: June 14, 2019
- Accepted Manuscript published: June 17, 2019 (version 1)
- Version of Record published: June 26, 2019 (version 2)
- Version of Record updated: July 1, 2019 (version 3)
Copyright
© 2019, Csanády & Töröcsik
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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