ATPase assay of purified, reconstituted CFTR protein.
The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a phosphorylation and nucleotide regulated chloride channel. CFTR also directly mediates the hydrolysis of ATP and this catalytic activity is loosely coupled to CFTR channel gating. However, mechanistic detail regarding the role of ATP hydrolysis in channel function is lacking. Our further understanding of the molecular basis for normal channel activity requires kinetic analysis of the ATPase activity by the full-length protein. This article describes an effective assay of ATPase activity by purified, reconstituted CFTR protein.
Kogan
Ilana
I
Department of Structural Biology and Biochemistry, The Hospital for Sick Children and Department of Physiology, University of Toronto, Ontario, Canada.
Ramjeesingh
Mohabir
M
Bear
Christine E
CE
D000251:Adenosine Triphosphatases / Q000378:metabolism* / Q000502:physiology; D019411:Clinical Laboratory Techniques; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D015640:Ion Channel Gating / Q000502:physiology*; D007700:Kinetics
C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000251:Adenosine Triphosphatases