Expression in Escherichia coli and characterisation of the human central CB1 and peripheral CB2 cannabinoid receptors |
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Authors: | Bernard Calandra Julie Tucker David Shire Reinhard Grisshammer |
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Affiliation: | (1) Sanofi Recherche, Centre de Labège, Labège-Innopole BP 137, 31676 Labège cedex, France;(2) Centre for Protein Engineering/MRC Centre, Hills Road, Cambridge, CB2 2QH, UK;(3) Centre for Protein Engineering/MRC Centre, Hills Road, Cambridge, CB2 2QH, UK |
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Abstract: | The human central CB1 and peripheral CB2 cannabinoid receptors were expressed in Escherichia coli as fusion proteins with the maltose-binding protein at their amino-termini and a hexa-histidine/Flag tag at their carboxyl-termini. Western blot analysis of the expressed proteins revealed considerable degradation of the CB1 fusion, which failed to bind either the cannabinoid agonist CP 55,940 or the CB1-specific antagonist SR 141716A. In contrast, the CB2 fusion was well-expressed and bound several cannabinoids with affinities comparable to those observed in mammalian expression systems. This revised version was published online in November 2006 with corrections to the Cover Date. |
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