Purification of functional full-length liver X receptor β produced in Escherichia coli

Gudrun Toresson, Gertrud U. Schuster, Knut R. Steffensen, Martin Bengtsson, Jan Ljunggren, Karin Dahlman-Wright, Jan Åke Gustafsson

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Liver X receptor β(LXRβ) is a ligand dependent transcription factor that is a member of the nuclear receptor superfamily. LXRβ and its isoform LXRα have recently been recognized as important regulators of lipid homeostasis in vertebrates. N-terminally hexahistidine-tagged rat LXRβ was expressed in Escherichia coli as a full-length protein and purified in two chromatographic steps, immobilized metal affinity chromatography and gel filtration. From 10 g of bacterial cells, 2.5 mg of protein was recovered. The purified LXRβ is functional with respect to ligand-, DNA-, and coactivator-binding. The synthetic ligand T0901317 bound to LXRβ with high affinity yielding a Kd of 2.7 nM. Specific interaction with DR4 response elements, in the presence of RXR, was demonstrated with electrophoretic mobility shift assay. Furthermore, surface plasmon resonance analysis of LXRβ binding to coactivator peptides revealed a ligand dependent interaction with the C-terminal nuclear receptor binding site of the coactivator RAP250. The purified LXRβ constitutes an important tool for further functional and structural studies.

Original languageEnglish (US)
Pages (from-to)190-198
Number of pages9
JournalProtein Expression and Purification
Issue number2
StatePublished - Jun 2004


  • Coactivator
  • Expression in E. coli
  • Hexahistidine tag
  • Liver X receptor
  • Nuclear receptor
  • RAP2050
  • Response element
  • Retinoid X receptor

ASJC Scopus subject areas

  • Biochemistry


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