Reconstitution of functional nuclear receptor proteins using high pressure refolding

Mol Genet Metab. 2005 Aug;85(4):318-22. doi: 10.1016/j.ymgme.2005.04.012.

Abstract

Members of the nuclear receptor superfamily are ligand dependent transcription factors and many of the receptors are difficult to produce in their functional form. Here, we describe a method for obtaining functional nuclear receptor ligand binding domain proteins from bacterial expressed inclusion bodies by high hydrostatic pressure induced refolding. High pressure refolding successfully reconstituted activity from several insoluble nuclear receptor proteins and represents a valuable tool for both functional and structural investigation of proteins or fragments thereof that might otherwise remain insoluble.

MeSH terms

  • Chromatography, Gel
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / physiology
  • Escherichia coli
  • Estrogen Receptor beta / chemistry
  • Estrogen Receptor beta / physiology
  • Humans
  • Hydrostatic Pressure
  • Inclusion Bodies
  • Plasmids / genetics
  • Protein Folding*
  • Receptors, Cytoplasmic and Nuclear / chemistry*
  • Receptors, Cytoplasmic and Nuclear / physiology
  • Transcription Factors / chemistry
  • Transcription Factors / physiology
  • Transcription, Genetic

Substances

  • DNA-Binding Proteins
  • Estrogen Receptor beta
  • NR5A2 protein, human
  • Receptors, Cytoplasmic and Nuclear
  • Transcription Factors
  • farnesoid X-activated receptor