Characterizing diverse orthologues of the cystic fibrosis transmembrane conductance regulator protein for structural studies

Biochem Soc Trans. 2015 Oct;43(5):894-900. doi: 10.1042/BST20150081.

Abstract

As an ion channel, the cystic fibrosis transmembrane conductance regulator (CFTR) protein occupies a unique niche within the ABC family. Orthologues of CFTR are extant throughout the animal kingdom from sharks to platypods to sheep, where the osmoregulatory function of the protein has been applied to differing lifestyles and diverse organ systems. In humans, loss-of-function mutations to CFTR cause the disease cystic fibrosis, which is a significant health burden in populations of white European descent. Orthologue screening has proved fruitful in the pursuit of high-resolution structural data for several membrane proteins, and we have applied some of the princples developed in previous studies to the expression and purification of CFTR. We have overexpressed this protein, along with evolutionarily diverse orthologues, in Saccharomyces cerevisiae and developed a purification to isolate it in quantities sufficient for structural and functional studies.

Keywords: CFTR; expression system; orthologues; purification.

Publication types

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

MeSH terms

  • Animals
  • Consensus Sequence
  • Cystic Fibrosis Transmembrane Conductance Regulator / chemistry*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / isolation & purification
  • Cystic Fibrosis Transmembrane Conductance Regulator / metabolism
  • Detergents / chemistry
  • Glycosylation
  • Humans
  • Models, Molecular*
  • Mutation
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / isolation & purification
  • Peptide Fragments / metabolism
  • Phosphorylation
  • Phylogeny
  • Protein Conformation
  • Protein Processing, Post-Translational*
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Solubility

Substances

  • CFTR protein, human
  • Detergents
  • Peptide Fragments
  • Recombinant Proteins
  • Cystic Fibrosis Transmembrane Conductance Regulator