The investigational Cystic Fibrosis drug Trimethylangelicin directly modulates CFTR by stabilizing the first membrane-spanning domain.
Cystic Fibrosis (CF) is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. The most common mutation, deletion of phenylalanine 508 (F508del), disrupts tertiary assembly, causing protein misprocessing and loss of CFTR function in epithelial tissues. Lumacaftor (VX-809) is a Class 1 corrector molecule shown to partially rescue misprocessing of F508del and together with the potentiator of channel activity: ivacaftor (VX-770) has been approved for treatment of CF patients homozygous for the F508del mutation. The specificity of these modulators for CFTR is thought to be conferred through direct binding. Trimethylangelicin (TMA) is a distinct small molecule modulator, previously shown to exhibit both corrector and potentiator activities. We were prompted to determine if TMA also mediates these activities by direct binding. Interestingly, we found that like VX-770, TMA was effective in enhancing anion efflux mediated by purified WT-CFTR reconstituted in phospholipid liposomes. Furthermore, like VX-809, TMA was effective in stabilizing the functional expression of CFTR lacking the regulatory "R" domain or second nucleotide-binding domain (NBD2). The smallest domain that was stabilized by TMA binding was the first membrane-spanning domain (MSD1) as previously observed for VX-809. Together, our findings support the claim that TMA binds directly to CFTR, and despite its distinct chemical structure, shares similar mechanisms as VX-770 and VX-809 to potentiate and stabilize CFTR, respectively.
Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Italy; Programme in Molecular Structure and Function, Hospital for Sick Children, Toronto, Canada.
Onofrio
O
Laselva
Programme in Molecular Structure and Function, Hospital for Sick Children, Toronto, Canada; Department of Biochemistry, University of Toronto, Canada.
Steven
S
Molinski
Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Italy.
Valeria
V
Casavola
Programme in Molecular Structure and Function, Hospital for Sick Children, Toronto, Canada; Department of Biochemistry, University of Toronto, Canada; Department of Physiology, University of Toronto, Canada. Electronic address: bear@sickkids.ca.
Christine E
CE
Bear
D000631:Aminopyridines / Q000737:chemistry / Q000494:pharmacology; D052117:Benzodioxoles / Q000737:chemistry / Q000494:pharmacology; D003550:Cystic Fibrosis; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D011564:Furocoumarins / Q000737:chemistry / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D015394:Molecular Structure; D000072417:Protein Domains
D000631:Aminopyridines; D052117:Benzodioxoles; D011564:Furocoumarins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C569105:lumacaftor; C041621:4,4',6-trimethylangelicin
Cystic Fibrosis; F508del; Modulator; Transmembrane domain; Trimethylangelicin; VX-809