1. Capturing the Direct Binding of CFTR Correctors to CFTR by Using Click Chemistry
- Author
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Sunitha Yarlagadda, Raymond A. Frizzell, Marcelo Actis, Chandrima Sinha, Koryse Woodroofe, Anjaparavanda P. Naren, Weiqiang Zhang, John P. Clancy, Naoaki Fujii, Songbai Lin, Kavisha Arora, Chang Suk Moon, and Assem G. Ziady
- Subjects
congenital, hereditary, and neonatal diseases and abnormalities ,Cystic Fibrosis ,Regulator ,Aminopyridines ,Cystic Fibrosis Transmembrane Conductance Regulator ,Plasma protein binding ,medicine.disease_cause ,Biochemistry ,Article ,Drug Discovery ,medicine ,Humans ,Benzodioxoles ,Molecular Biology ,Mutation ,biology ,Chemistry ,Drug discovery ,Organic Chemistry ,HEK 293 cells ,Transmembrane protein ,Cystic fibrosis transmembrane conductance regulator ,Cell biology ,HEK293 Cells ,Mechanism of action ,biology.protein ,Molecular Medicine ,Click Chemistry ,medicine.symptom ,Protein Binding - 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 Cu(I) -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.
- Published
- 2015