title: Effects of co-expression of liver X receptor β-ligand binding domain with its partner, retinoid X receptor α-ligand binding domain, on their solubility and biological activity in Escherichia coli.
abstract: In this presentation, I describe the expression and purification of the recombinant liver X receptor β-ligand binding domain proteins in E. coli using a commercially available double cistronic vector, pACYCDuet-1, to express the receptor heterodimer in a single cell as the soluble form. I describe here the expression and characterization of a biologically active heterodimer composed of the liver X receptor β-ligand binding domain and retinoid X receptor α-ligand binding domain. Although many of these proteins were previously seen to be produced in E. coli as insoluble aggregates or "inclusion bodies", I show here that as a form of heterodimer they can be made in soluble forms that are biologically active. This suggests that co-expression of the liver X receptor β-ligand binding domain with its binding partner improves the solubility of the complex and probably assists in their correct folding, thereby functioning as a type of molecular chaperone.
authors.affiliation: Department of Medical Laboratory Science, College of Health Science, Dankook University, Cheonan-si 330-714, Republic of Korea.
authors.forename: Hyun
authors.initials: H
authors.lastname: Kang
mesh_terms: D001665:Binding Sites; D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D002479:Inclusion Bodies / Q000378:metabolism / Q000382:microbiology; D008024:Ligands; D000071518:Liver X Receptors; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D057093:Orphan Nuclear Receptors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D047490:Retinoid X Receptor alpha / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D014157:Transcription Factors
chemical_list: D008024:Ligands; D000071518:Liver X Receptors; D018832:Molecular Chaperones; D057093:Orphan Nuclear Receptors; D011994:Recombinant Proteins; D047490:Retinoid X Receptor alpha; D014157:Transcription Factors