Purification and crystallization of the cystic fibrosis transmembrane conductance regulator (CFTR)

J Biol Chem. 2004 Sep 10;279(37):39051-7. doi: 10.1074/jbc.M407434200. Epub 2004 Jul 9.

Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) is a membrane protein that is mutated in patients suffering from cystic fibrosis. Here we report the purification and first crystallization of wild-type human CFTR. Functional characterization of the material showed it to be highly active. Electron crystallography of negatively stained two-dimensional crystals of CFTR has revealed the overall architecture of this channel for two different conformational states. These show a strong structural homology to two conformational states of another eukaryotic ATP-binding cassette transporter, P-glycoprotein. In contrast to P-glycoprotein, however, both conformational states can be observed in the presence of a nucleotide, which may be related to the role of CFTR as an ion channel rather than a transporter. The hypothesis that the two conformations could represent the "open" and "closed" states of the channel is considered.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / chemistry
  • ATP-Binding Cassette Transporters / chemistry
  • Adenosine Triphosphate / chemistry
  • Animals
  • Bacterial Proteins / chemistry
  • Cricetinae
  • Crystallography, X-Ray
  • Cystic Fibrosis Transmembrane Conductance Regulator / chemistry*
  • Cystic Fibrosis Transmembrane Conductance Regulator / isolation & purification*
  • Dimerization
  • Dose-Response Relationship, Drug
  • Humans
  • Hydrolysis
  • Kidney / metabolism
  • Microscopy, Electron
  • Nucleotides / chemistry
  • Protein Conformation

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • ATP-Binding Cassette Transporters
  • Bacterial Proteins
  • CFTR protein, human
  • MsbA protein, Bacteria
  • Nucleotides
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Adenosine Triphosphate