Expression and purification of CB2 for NMR studies in micellar solution.
We demonstrate feasibility of biophysical characterization of the peripheral cannabinoid receptor CB2 produced by heterologous expression in E. coli membranes. Recombinant receptor was purified by affinity chromatography, and NMR diffusion experiments performed on CB2 solubilized in dodecylphosphocholine (DPC) micelles. Circular dichroism spectroscopy indicated high alpha-helical content (49 %) of CB2.
Krepkiy
Dmitriy
D
National Institute on Alcoholism and Alcohol Abuse, National Institutes of Health, Bethesda, MD 20892-9410, USA.
Gawrisch
Klaus
K
Yeliseev
Alexei
A
D002186:Cannabinoids / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics; D002942:Circular Dichroism; D003902:Detergents / Q000737:chemistry; D004058:Diffusion; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D058619:Maltose-Binding Proteins; D008823:Micelles; D043885:Receptor, Cannabinoid, CB2 / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012996:Solutions
D002186:Cannabinoids; D002352:Carrier Proteins; D003902:Detergents; D044385:GTP-Binding Protein alpha Subunits; D008024:Ligands; D058619:Maltose-Binding Proteins; D008823:Micelles; D043885:Receptor, Cannabinoid, CB2; D011993:Recombinant Fusion Proteins; D012996:Solutions