Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / metabolism
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Animals
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Caprylates
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Cell Line
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Chlorides / metabolism
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Chromatography, Affinity
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Chromatography, Thin Layer
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Cloning, Molecular
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Cyclic AMP-Dependent Protein Kinases / metabolism
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Cystic Fibrosis Transmembrane Conductance Regulator / genetics
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Cystic Fibrosis Transmembrane Conductance Regulator / isolation & purification*
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Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
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Detergents
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Fluorocarbons
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Humans
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Ion Channel Gating
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Liposomes / metabolism
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Oligodeoxyribonucleotides / chemistry
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Phosphorylation
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Spodoptera / genetics
Substances
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CFTR protein, human
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Caprylates
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Chlorides
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Detergents
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Fluorocarbons
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Liposomes
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Oligodeoxyribonucleotides
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Recombinant Proteins
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Cystic Fibrosis Transmembrane Conductance Regulator
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Adenosine Triphosphate
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perfluorooctanoic acid
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Cyclic AMP-Dependent Protein Kinases
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Adenosine Triphosphatases