title: S-Nitrosothiols increases cystic fibrosis transmembrane regulator expression and maturation in the cell surface.
abstract: S-nitrosothiols (SNOs) are endogenous signaling molecules with a broad spectrum of beneficial airway effects. SNOs are normally present in the airway, but levels tend to be low in cystic fibrosis (CF) patients. We and others have demonstrated that S-nitrosoglutathione (GSNO) increases the expression, maturation, and function of wild-type and mutant F508del cystic fibrosis transmembrane conductance regulator (CFTR) in human bronchial airway epithelial (HBAE) cells. We hypothesized that membrane permeable SNOs, such as S-nitrosoglutathione diethyl ester (GNODE) and S-nitroso-N-acetyl cysteine (SNOAC) may be more efficient in increasing the maturation of CFTR. HBAE cells expressing F508del CFTR were exposed to GNODE and SNOAC. The effects of these SNOs on the expression and maturation of F508del CFTR were determined by cell surface biotinylation and Western blot analysis. We also found for the first time that GNODE and SNOAC were effective at increasing CFTR maturation at the cell surface. Furthermore, we found that cells maintained at low temperature increased cell surface stability of F508del CFTR whereas the combination of low temperature and SNO treatment significantly extended the half-life of CFTR. Finally, we showed that SNO decreased the internalization rate of F508del CFTR in HBAE cells. We anticipate identifying the novel mechanisms, optimal SNOs, and lowest effective doses which could benefit cystic fibrosis patients.
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA. Electronic address: kxz91@case.edu.
authors.forename: Khalequz
authors.initials: K
authors.lastname: Zaman
authors.affiliation: Pediatric Respiratory Medicine, Department of Pediatrics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
authors.forename: Deric
authors.initials: D
authors.lastname: Bennett
authors.affiliation: Pediatric Respiratory Medicine, Department of Pediatrics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
authors.forename: Maya
authors.initials: M
authors.lastname: Fraser-Butler
authors.affiliation: Pediatric Respiratory Medicine, Department of Pediatrics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
authors.forename: Zivi
authors.initials: Z
authors.lastname: Greenberg
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Paulina
authors.initials: P
authors.lastname: Getsy
authors.affiliation: Epidemiology and Biostatistics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Abdus
authors.initials: A
authors.lastname: Sattar
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Laura
authors.initials: L
authors.lastname: Smith
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Deborah
authors.initials: D
authors.lastname: Corey
authors.affiliation: Department of Physiology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
authors.affiliation: Pediatric Respiratory Medicine, Department of Pediatrics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA.
authors.forename: John
authors.initials: J
authors.lastname: Hunt
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Lewis
authors.affiliation: Pediatric Pulmonology Division, Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gaston
mesh_terms: D001980:Bronchi / Q000187:drug effects / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics* / Q000378:metabolism*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D026403:S-Nitrosothiols / Q000378:metabolism / Q000494:pharmacology*; D017384:Sequence Deletion; D015398:Signal Transduction
chemical_list: C505032:CFTR protein, human; D050505:Mutant Proteins; D026403:S-Nitrosothiols; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFTR; Chaperones; Cystic fibrosis; Molecular therapy; S-nitrosothiol; S-nitrosylation