Protein solubility and folding monitored in vivo by structural complementation of a genetic marker protein

Nat Biotechnol. 2001 Feb;19(2):131-6. doi: 10.1038/84389.

Abstract

Protein misfolding is the basis of a number of human diseases and presents an obstacle to the production of soluble recombinant proteins. We present a general method to assess the solubility and folding of proteins in vivo. The basis of this assay is structural complementation between the alpha- and omega- fragments of beta-galactosidase (beta-gal). Fusions of the alpha-fragment to the C terminus of target proteins with widely varying in vivo folding yield and/or solubility levels, including the Alzheimer's amyloid beta (A beta) peptide and a non-amyloidogenic mutant thereof, reveal an unambiguous correlation between beta-gal activity and the solubility/folding of the target. Thus, structural complementation provides a means of monitoring protein solubility/misfolding in vivo, and should find utility in the screening for compounds that influence the pathological consequences of these processes.

Publication types

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

MeSH terms

  • Amyloid beta-Peptides / chemistry*
  • Amyloid beta-Peptides / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator / chemistry*
  • Cystic Fibrosis Transmembrane Conductance Regulator / genetics
  • Genes, Reporter
  • Genetic Markers
  • Humans
  • Mutagenesis, Site-Directed
  • Peptide Fragments / chemistry
  • Protein Folding*
  • Recombinant Fusion Proteins / chemistry*
  • Solubility
  • beta-Galactosidase / chemistry*
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism

Substances

  • Amyloid beta-Peptides
  • CFTR protein, human
  • Genetic Markers
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • beta-Galactosidase