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Recombinant Expression and Purification of Cannabinoid Receptor CB2, a G Protein-Coupled Receptor
1National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, MD, USA. yeliseeva@mail.nih.gov.
Methods in Molecular Biology (Clifton, N.J.)
|June 4, 2021
Summary
Researchers purified the human cannabinoid receptor 2 (CB2), a G protein-coupled receptor (GPCR), using E. coli and mammalian cells. This method yields milligram quantities for structural and functional studies.
Area of Science:
- Biochemistry
- Structural Biology
- Pharmacology
Background:
- G protein-coupled receptors (GPCRs) are crucial membrane proteins regulating cellular functions.
- Understanding GPCR structure and activity requires pure, stable, milligram-scale protein preparations.
- The human cannabinoid receptor 2 (CB2) is a key GPCR involved in various physiological processes.
Purpose of the Study:
- To develop a robust method for expressing and purifying the type II human cannabinoid receptor CB2.
- To obtain milligram quantities of purified CB2 receptor suitable for downstream applications.
- To compare expression and purification yields in different host systems.
Main Methods:
- Expression of type II human cannabinoid receptor CB2 in Escherichia coli.
- Expression of type II human cannabinoid receptor CB2 in Expi293 mammalian suspension cells.
- Purification of recombinant CB2 receptor using affinity chromatography and other standard protein purification techniques.
Main Results:
- Successful expression and purification of milligram quantities of the human CB2 receptor.
- Demonstrated suitability of the purified receptor for structural and functional analyses.
- Comparison of receptor yield and purity between E. coli and Expi293 expression systems.
Conclusions:
- The described method enables the production of high-purity, milligram-scale human CB2 receptor.
- This provides a valuable resource for detailed structural and functional investigations of CB2.
- The findings facilitate further research into GPCRs and their therapeutic potential.
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