Biochemical evidence for ATPase activity in CFTR-enriched apical membrane vesicles from tracheal epithelium.
In apical membrane vesicles from beef tracheal epithelia expressing up to 30% of the proteins as functional cystic fibrosis transmembrane conductance regulator (CFTR)-- i.e. a voltage-independent and PKA-sensitive 36Cl- flux--an ATPase activity, different from P, F0F1 and V types, was reproducibly detected. Its specific activity averaged 20 micromol Pi h(-1) mg(-1) with an apparent affinity for ATP of 530 +/- 30 microM. Its possible involvement in CFTR functions was supported by (1) the linear relationship between the ATPase activity and the magnitude of 36Cl- fluxes (turnover rate: 3 ATP hydrolyzed per CFTR per second), (2) the same rank of potency of ATP, ITP, GTP, UTP and CTP to be hydrolyzed and to open CFTR chloride channels, (3) the similar and parallel inhibition of the ATPase and CFTR Cl- fluxes by NS004 (IC50: 60 microM) and (4) the potency of anti-R domain antibodies to increase by 18% the ATPase activity.
Laboratoire de Pharmacologie des Transports Ioniques Membranaires, Université Paris 7-Denis Diderot, France.
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D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001562:Benzimidazoles / Q000494:pharmacology; D002417:Cattle; D002712:Chlorides / Q000378:metabolism; D002733:Chlorophenols / Q000494:pharmacology; D018699:Coated Vesicles / Q000201:enzymology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000032:analysis / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000201:enzymology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D007700:Kinetics; D011868:Radioisotopes; D013379:Substrate Specificity; D014132:Trachea / Q000201:enzymology*
D000906:Antibodies; D001562:Benzimidazoles; D002712:Chlorides; D002733:Chlorophenols; D004791:Enzyme Inhibitors; D011868:Radioisotopes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C083230:NS 004; D000255:Adenosine Triphosphate; D000469:Alkaline Phosphatase; D000251:Adenosine Triphosphatases