A novel procedure for the efficient purification of the cystic fibrosis transmembrane conductance regulator (CFTR).
This report describes a novel, single-step strategy for the purification of the cystic fibrosis transmembrane conductance regulator from Sf9 cells, which will facilitate studies of the structure-function relationships of this clinically important molecule. The new method combines the use of the novel detergent sodium pentadecafluoro-octanoate with metal-affinity chromatography to produce a high yield of purified protein which can be functionally reconstituted as a chloride channel and an ATPase.
Ramjeesingh
M
M
Division of Cell Biology, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Canada M5G 1X8.
Li
C
C
Garami
E
E
Huan
L J
LJ
Hewryk
M
M
Wang
Y
Y
Galley
K
K
Bear
C E
CE
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002210:Caprylates; D002460:Cell Line; D002712:Chlorides / Q000378:metabolism; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents; D005466:Fluorocarbons; D006801:Humans; D015640:Ion Channel Gating; D008081:Liposomes / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics
C505032:CFTR protein, human; D002210:Caprylates; D002712:Chlorides; D003902:Detergents; D005466:Fluorocarbons; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000255:Adenosine Triphosphate; C023036:perfluorooctanoic acid; D017868:Cyclic AMP-Dependent Protein Kinases; D000251:Adenosine Triphosphatases