A novel procedure for the efficient purification of the cystic fibrosis transmembrane conductance regulator (CFTR)

Biochem J. 1997 Oct 1;327 ( Pt 1)(Pt 1):17-21. doi: 10.1042/bj3270017.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Adenosine Triphosphate / metabolism
  • Animals
  • Caprylates
  • Cell Line
  • Chlorides / metabolism
  • Chromatography, Affinity
  • Chromatography, Thin Layer
  • Cloning, Molecular
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / isolation & purification*
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • Detergents
  • Fluorocarbons
  • Humans
  • Ion Channel Gating
  • Liposomes / metabolism
  • Oligodeoxyribonucleotides / chemistry
  • Phosphorylation
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Spodoptera / genetics

Substances

  • CFTR protein, human
  • Caprylates
  • Chlorides
  • Detergents
  • Fluorocarbons
  • Liposomes
  • Oligodeoxyribonucleotides
  • Recombinant Proteins
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Adenosine Triphosphate
  • perfluorooctanoic acid
  • Cyclic AMP-Dependent Protein Kinases
  • Adenosine Triphosphatases