Isolation and purification of morphine receptor by affinity chromatography.
Brain membranes were solubilized by sonication and Triton X-100 extraction and applied to an affinity column consisting of a 6-succinyl morphine derivative of Affi Gel-102. A fraction exhibiting high opiate binding was eluted by tris-buffer containing naloxone, CHAPS and NaCl. This fraction consisted of both proteins and acidic lipids. The opiate binding properties of this purified material exhibited many properties similar to those of membrane bound receptors of the u-type, including high affinity, stereospecificity, Na-effect and rank order in affinity for opiates. This opiate binding material was highly sensitive to both trypsin and N-ethylmaleimide. Based on the protein content of the isolated membrane receptor, a 3200-fold purification over the original brain P2 fraction was achieved.
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T M
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D000818:Animals; D001667:Binding, Competitive; D001923:Brain Chemistry; D002846:Chromatography, Affinity; D005033:Ethylmaleimide; D006160:Guanosine Triphosphate; D008563:Membrane Lipids / Q000302:isolation & purification / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols; D051381:Rats; D011957:Receptors, Opioid / Q000302:isolation & purification* / Q000378:metabolism; D012965:Sodium Chloride; D013010:Sonication; D014357:Trypsin
D008563:Membrane Lipids; D009419:Nerve Tissue Proteins; D011957:Receptors, Opioid; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D006160:Guanosine Triphosphate; D017830:Octoxynol; D014357:Trypsin; D005033:Ethylmaleimide