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Alexei A Yeliseev

Alexei A Yeliseev

National Institute on Alcoholism and Alcohol Abuse USA

Title: Human cannabinoid receptor CB2: Expression, functional and structural studies

Biography

Biography: Alexei A Yeliseev

Abstract

The human cannabinoid receptor CB2 belongs to the class A of hepta helical G protein-coupled receptors (GPCR) and is an attractive target for the development of drugs for management of pain, inflammation, osteoporosis and treatment of immunological disorders. High resolution structural studies are critical to obtain insights into the molecular mechanisms of ligand binding and activation of CB2. We developed methods for expression in milligram quantities, purification, reconstitution in lipid bilayers and stabilization of the functional recombinant CB2 as well as efficient stable isotope labeling of CB2 by high density fermentation were developed enabling NMR studies. NMR analysis of labeled receptor reconstituted in proteoliposomes in agonist or inverse-agonist-bound form will be reported. Monoclonal antibodies were raised against the purified CB2 and the affinity of interaction with CB2 was determined by surface plasmon resonance. Finally, we demonstrate the specific effects of lipids with negatively charged head group in activation of CB2 reconstituted in proteoliposomes as measured by an in vitro G protein activation which may have important physiological significance as manifested in natural membranes of various lipid compositions.