Interdomain but not intermolecular interactions observed in CFTR channels.
Gating of the cystic fibrosis transmembrane conductance regulator (CFTR) channels requires interdomain and/or intermolecular interactions involving different parts of the protein, yet the exact nature of those interactions remains unclear. In this study we report that treating wild type CFTR-expressing cells with oxidizing agents results in a significant reduction in the gel mobility of the protein indicative of the formation of disulfide bonds. In contrast, mutant CFTR channels in which cysteine residues in both nucleotide binding domains (NBDs) were mutated to serine, showed little change in gel mobility in oxidizing conditions. Mutation of the two cysteine residues in either the first or the second NBD alone also eliminates the change in gel mobility in oxidizing conditions. Wild type channels treated with oxidizing agents did not appear to form disulfide bonds with other proteins, suggesting that the close association that allows the formation of disulfide bonds occurs only within single proteins and not between separate channels interacting in a multimer.
Kembi
F
F
Department of Biology, Delaware State University, 1200 DuPont Highway, Dover, Delaware 19901, USA.
Harrington
M A
MA
D019943:Amino Acid Substitution / Q000235:genetics; D001665:Binding Sites; D002460:Cell Line; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D015640:Ion Channel Gating; D008623:Mercaptoethanol / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D009711:Nucleotides / Q000378:metabolism; D016877:Oxidants / Q000378:metabolism; D010084:Oxidation-Reduction; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D014162:Transfection
C505032:CFTR protein, human; D004220:Disulfides; D009711:Nucleotides; D016877:Oxidants; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D008623:Mercaptoethanol; D006861:Hydrogen Peroxide; D003545:Cysteine