title: Capturing the Direct Binding of CFTR Correctors to CFTR by Using Click Chemistry.
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.
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.|Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, 38163, USA.
authors.forename: Chandrima
authors.initials: C
authors.lastname: Sinha
authors.affiliation: Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, 38163, USA.|Department of Pediatrics, University of Tennessee Health Science Center, Memphis, TN, 38103, USA.
authors.forename: Weiqiang
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Chang Suk
authors.initials: CS
authors.lastname: Moon
authors.affiliation: Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, 38105, USA.
authors.forename: Marcelo
authors.initials: M
authors.lastname: Actis
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Sunitha
authors.initials: S
authors.lastname: Yarlagadda
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Kavisha
authors.initials: K
authors.lastname: Arora
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Koryse
authors.initials: K
authors.lastname: Woodroofe
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: John P
authors.initials: JP
authors.lastname: Clancy
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Songbai
authors.initials: S
authors.lastname: Lin
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA.
authors.forename: Assem G
authors.initials: AG
authors.lastname: Ziady
authors.affiliation: Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA.
authors.forename: Raymond
authors.initials: R
authors.lastname: Frizzell
authors.affiliation: Department of Chemical Biology and Therapeutics, St. Jude Children's Research Hospital, Memphis, TN, 38105, USA.
authors.forename: Naoaki
authors.initials: N
authors.lastname: Fujii
authors.affiliation: Division of Pulmonary Medicine, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, 45229, USA. anaren@cchmc.org.|Department of Physiology, University of Tennessee Health Science Center, Memphis, TN, 38163, USA. anaren@cchmc.org.
authors.forename: Anjaparavanda P
authors.initials: AP
authors.lastname: Naren
mesh_terms: D000631:Aminopyridines / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D052117:Benzodioxoles / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D057930:Click Chemistry; D003550:Cystic Fibrosis / Q000188:drug therapy / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism*; D055808:Drug Discovery; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D011485:Protein Binding
chemical_list: D000631:Aminopyridines; D052117:Benzodioxoles; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C569105:lumacaftor
keywords: CFTR correctors; VX-809; click chemistry; cystic fibrosis; protein expression