Generating Proteins for High-Throughput Drug Screening Using Customized Plasmids for Expressing Nuclear Receptor Proteins in E. Coli

SLU ID 19-012 | Customized Plasmids That Express Stable Ligand Binding Domains for TLX, ERRalpha, ERRbeta, ERRgamma, REVERBalpha, and REVERBbeta in E. Coli Cells

Background

There are currently 48 known nuclear receptors in the human genome, each containing an NH2 and a COOH tail with a transcriptional regulation domain, a DNA binding domain, and a ligand binding domain in between. The functions of all 48 known nuclear receptors are not known; those without known actions are called orphan receptors. PPAR receptors are involved in metabolic regulation and are important drug targets. A current problem with nuclear receptors lies in establishing a high yield of stable ligand binding domains without suffering protein unfolding.

Overview

Researchers at Saint Louis University have designed plasmids that express stable ligand binding domains of TLX, ERRalpha, ERRbeta, ERRgamma, REVERBalpha, and REVERBbeta in Escherichia coli (E. coli) cells. They have also optimized protein expression and purification protocols for these receptors.

Benefits

The potential benefits of this technology include:

• Increasing protein stability

• Increasing protein yield

• Increasing the temperature range for enzymatic cleavage

• Minimizing protein unfolding

• Increasing buffer conditions optimization

Applications

Potential applications of this technology include:

• High throughput drug screening

• Biophysical assays

• Structure elucidation

• Protein expression

• Protein purification

Opportunity

Saint Louis University is seeking partners to further develop and commercialize this technology.