PLoS ONE 2013-01-01

Expression and purification of functional human mu opioid receptor from E.coli.

Yanbin Ma, Jan Kubicek, Jörg Labahn

Index: PLoS ONE 8 , e56500, (2013)

Full Text: HTML

Abstract

N-terminally his-tagged human mu opioid receptor, a G protein-coupled receptor was produced in E.coli employing synthetic codon-usage optimized constructs. The receptor was expressed in inclusion bodies and membrane-inserted in different E.coli strains. By optimizing the expression conditions the expression level for the membrane-integrated receptor was raised to 0.3-0.5 mg per liter of culture. Milligram quantities of receptor could be enriched by affinity chromatography from IPTG induced cultures grown at 18°C. By size exclusion chromatography the protein fraction with the fraction of alpha-helical secondary structure expected for a 7-TM receptor was isolated, by CD-spectroscopy an alpha-helical content of ca. 45% was found for protein solubilised in the detergent Fos-12. Receptor in Fos-12 micelles was shown to bind endomorphin-1 with a K(D) of 61 nM. A final yield of 0.17 mg functional protein per liter of culture was obtained.

Related Compounds

  • Spautin-1

Related Articles:

Single Chain Variable Fragments Produced in Escherichia coli against Heat-Labile and Heat-Stable Toxins from Enterotoxigenic E. coli.

2015-01-01

[PLoS ONE 10 , e0131484, (2015)]

Photosynthetic production of enantioselective biocatalysts.

2015-01-01

[Microb. Cell Fact. 14 , 53, (2015)]

Outer-membrane translocation of bulky small molecules by passive diffusion.

2015-06-09

[Proc. Natl. Acad. Sci. U. S. A. 112 , E2991-9, (2015)]

Effects of pre- and postnatal exposure to flutamide on connexin 43 expression in testes and ovaries of prepubertal pigs.

2010-01-01

[Eur. J. Histochem. 54(2) , e15, (2010)]

Cellulose Microfibril Formation by Surface-Tethered Cellulose Synthase Enzymes.

2016-02-23

[ACS Nano 10 , 1896-907, (2016)]

More Articles...