Reconstitution of functional nuclear receptor proteins using high pressure refolding.
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.
Schoner
Brigitte E
BE
Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285, USA.
Bramlett
Kelli S
KS
Guo
Haihong
H
Burris
Thomas P
TP
D002850:Chromatography, Gel; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000502:physiology; D004926:Escherichia coli; D047629:Estrogen Receptor beta / Q000737:chemistry / Q000502:physiology; D006801:Humans; D006874:Hydrostatic Pressure; D002479:Inclusion Bodies; D010957:Plasmids / Q000235:genetics; D017510:Protein Folding; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000502:physiology; D014157:Transcription Factors / Q000737:chemistry / Q000502:physiology; D014158:Transcription, Genetic; D000099150:Receptor, Farnesoid X-Activated
D004268:DNA-Binding Proteins; D047629:Estrogen Receptor beta; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D000099150:Receptor, Farnesoid X-Activated; C082631:NR5A2 protein, human