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Protein Expr Purif. 2004 Feb;33(2):176-84.

Purification and characterization of the recombinant human dopamine D2S receptor from Pichia pastoris.

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  • 1Department of Bioanalysis and Toxicology, University Centre for Pharmacy, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.


The human dopamine D2S receptor was expressed in the methylotrophic yeast Pichia pastoris, where the receptor with a molecular mass of approximately 40kDa exhibited specific and saturable binding properties. The dopamine antagonist [3H]spiperone showed an average dissociation constant K(d) of 0.6+/-0.17 nM for the dopamine D2S receptor. The receptor was solubilized using the non-ionic detergent dodecylmaltoside and purified by affinity chromatography using a Ni(2+) chelate (His-Trap) column or by batch extraction with an anti-FLAG M1 affinity resin. The receptor maintained its biological activity after solubilization and purification from the membrane protein fraction. A 244- or 185-fold enrichment, as judged by an increase in specific binding, was obtained after adsorption to the His-Trap or anti-FLAG materials, respectively.

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