Solubilization in high yield of opioid receptors retaining high-affinity delta, mu and kappa binding sites |
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Authors: | C D Demoliou-Mason E A Barnard |
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Institution: | 1. Dipartimento di Fisica, Universita'' della Calabria, Arcacavata, Italy;2. Laboratorium für Biochemie I, Eidgenoessische Technische Hochschule ETH Zentrum, Universitätsstrasse 16, 8092 Zurich, Switzerland |
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Abstract: | The binding sites for opiates (agonist and antagonist) and opioid peptides can be solubilized from rat brain membranes with digitonin in the presence of Mg2+ (10 mM). High affinity and high capacity binding to the soluble delta, mu, and kappa receptors is obtainable when the membranes are treated in Mg2+ (30 degrees C, 60 min) prior to solubilization. The yields of solubilized binding sites extracted with digitonin, 40-90%, are higher than those obtained from Mg2+-pretreated membranes with other detergents commonly used for receptor solubilization. The stability of the digitonin-soluble opioid receptor at room temperature makes it useful for purification and characterization. |
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Keywords: | ERP Hemoglobin Liver fluke Distal histidine |
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