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

W. Christian Wigley, Rhesa D. Stidham, Nathan M. Smith, John F. Hunt, Philip J. Thomas

Research output: Contribution to journalArticle

132 Scopus citations

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 α- and ω-fragments of β-galactosidase (β-gal). Fusions of the α-fragment to the C terminus of target proteins with widely varying in vivo folding yield and/or solubility levels, including the Alzheimer's amyloid β (Aβ) peptide and a non-amyloidogenic mutant thereof, reveal an unambiguous correlation between β-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.

Original languageEnglish (US)
Pages (from-to)131-136
Number of pages6
JournalNature biotechnology
Volume19
Issue number2
DOIs
StatePublished - Feb 21 2001

Keywords

  • Aggregation
  • Complementation
  • Folding
  • Protein-folding diseases
  • Solubility

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Fingerprint Dive into the research topics of 'Protein solubility and folding monitored in vivo structural complementation of a genetic marker protein'. Together they form a unique fingerprint.

  • Cite this