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Capturing the Direct Binding of CFTR Correctors to CFTR by Using Click Chemistry |
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| Authors: | Dr Chandrima Sinha Prof Weiqiang Zhang Dr Chang Suk Moon Marcelo Actis Sunitha Yarlagadda Dr Kavisha Arora Dr Koryse Woodroofe Prof John P Clancy Dr Songbai Lin Prof Assem G Ziady Prof Raymond Frizzell Prof Naoaki Fujii Prof Anjaparavanda P Naren |
| Affiliation: | 1. Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA;2. Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, USA;3. Department of Pediatrics, University of Tennessee Health Science Center, Memphis, TN, USA;4. Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, USA;5. Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA |
| Abstract: | Cystic fibrosis (CF) is a lethal genetic disease caused by the loss or dysfunction of the CF transmembrane conductance regulator (CFTR) channel. F508del is the most prevalent mutation of the CFTR gene and encodes a protein defective in folding and processing. VX‐809 has been reported to facilitate the folding and trafficking of F508del‐CFTR and augment its channel function. The mechanism of action of VX‐809 has been poorly understood. In this study, we sought to answer a fundamental question underlying the mechanism of VX‐809: does it bind CFTR directly in order to exert its action? We synthesized two VX‐809 derivatives, ALK‐809 and SUL‐809, that possess an alkyne group and retain the rescue capacity of VX‐809. By using CuI‐catalyzed click chemistry, we provide evidence that the VX‐809 derivatives bind CFTR directly in vitro and in cells. Our findings will contribute to the elucidation of the mechanism of action of CFTR correctors and the design of more potent therapeutics to combat CF. |
| Keywords: | CFTR correctors click chemistry cystic fibrosis protein expression VX-809 |