title: Cystic fibrosis transmembrane conductance regulator in human and mouse red blood cell membranes and its interaction with ecto-apyrase.
abstract: Elevated blood ATP and increased red blood cell (RBC) ATP transport is associated with cystic fibrosis (CF). In this report, we demonstrate the presence of the wild-type and the DeltaF508 mutant form of the CF transmembrane conductance regulator protein in RBC membranes and its putative interaction with ecto-apyrase, an ATP hydrolyzing enzyme also present in the RBC membrane. RBC membranes of control and DeltaF508 individuals and of wild-type and CF transmembrane conductance regulator-knockout mice were examined by immunoblot using several antibodies directed against different epitopes of this protein. These experiments indicated that human RBC membranes contain comparable amounts of the wild-type CF transmembrane conductance regulator protein and the DeltaF508 mutant form of the protein, respectively. CF transmembrane conductance regulator protein was also detected in wild-type mouse RBC membranes but not in the gene knockout mouse RBC membranes. Antibodies directed against ecto-apyrase co-immunoprecipitated CF transmembrane conductance regulator protein of human RBC membranes indicating a physical interaction between these two membrane proteins consistent with ATP transport and extracellular hydrolysis. We conclude that RBCs are a significant repository of CF transmembrane conductance regulator protein and should provide a novel system for evaluating its expression and function.
authors.affiliation: Dartmouth Medical School, Lebanon, New Hampshire, USA.
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Sterling
authors.forename: Sanjiv
authors.initials: S
authors.lastname: Shah
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Kim
authors.forename: Nicholas I F
authors.initials: NI
authors.lastname: Johnston
authors.forename: Anna Y
authors.initials: AY
authors.lastname: Salikhova
authors.forename: Edward H
authors.initials: EH
authors.lastname: Abraham
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D015703:Antigens, CD; D001081:Apyrase / Q000378:metabolism*; D001692:Biological Transport / Q000502:physiology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D011485:Protein Binding / Q000502:physiology
chemical_list: D015703:Antigens, CD; C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000255:Adenosine Triphosphate; D001081:Apyrase; C067869:CD39 antigen