search
for
 About Bioline  All Journals  Testimonials  Membership  News  Donations


Tropical Journal of Pharmaceutical Research
Pharmacotherapy Group, Faculty of Pharmacy, University of Benin, Benin City, Nigeria
ISSN: 1596-5996
EISSN: 1596-5996
Vol. 13, No. 8, 2014, pp. 1247-1255
Bioline Code: pr14172
Full paper language: English
Document type: Research Article
Document available free of charge

Tropical Journal of Pharmaceutical Research, Vol. 13, No. 8, 2014, pp. 1247-1255

 en Biological Activities of Recombinant Liver X Receptor β- Ligand Binding Domain Protein in Tetracycline-Inducible Expression System
Kang, Hyun

Abstract

Purpose: To investigate tetracycline-inducible expression system for producing clinically usable, highquality liver X receptor ligand-binding domain recombinant protein.
Methods: In this study, we have expressed and purified the recombinant liver X receptor β-ligand binding domain proteins in E. coli check for this species in other resources using a tetracycline inducible system. To allow for biological activities, we subcloned into pPROTet.E HN vector, expressed in E. coli cells under optimized conditions, purified and characterized the recombinant liver X receptor β-ligand-binding domain proteins using fluorescence polarization assay.
Results: The use of pPROTet.E HN vector simplified downstream purification processes, including cleavage and elution thereby increasing the solubility and yield of the protein of interest. There was a 2.3-fold increase in the efficiency of recombinant LXR β-ligand binding domain (LBD) production by optimizing the expression temperature to 15 °C when compared to those induced at 37°C during the induction procedures. A typical dose-response curve obtained using increasing concentrations of the purified recombinant LXR β-LBD (197-461) and measuring fluorescence intensity (FI) as an index of fluorescent peptide binding to LBD showed 50 % effective dose (ED50) value of 533 nM. The recombinant LXR β-LBDs were substantially soluble and should be useful for future biological, biophysical and structural analyses of nuclear receptor complexes. This may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.
Conclusion: These findings indicate that recombinant LXR β-LBD protein is a promising target for the development of molecular ligands with improved therapeutic windows.

Keywords
Nuclear receptor; Recombinant LXR β-LBD; Tetracycline-inducible expression system; Fluorescence polarization assay

 
© Copyright 2014 - Tropical Journal of Pharmaceutical Research
Alternative site location: http://www.tjpr.org

Home Faq Resources Email Bioline
© Bioline International, 1989 - 2022, Site last up-dated on 27-Jul-2022.
Site created and maintained by the Reference Center on Environmental Information, CRIA, Brazil
System hosted by the Internet Data Center of Rede Nacional de Ensino e Pesquisa, RNP, Brazil