Chromatography and partial purification of solubilized opiate receptors.
Sheep brain membrane opiate receptors were solubilized using two different approaches: (A) Chaps was used to solubilize [3H]buprenorphine labeled membranes. Macromolecular labeled material had a stokes radius of approximately 90 A. Treatment with phospholipase A2, but not phospholipase C, decreased the stokes radius to 50 A indicating the presence and eventual structural role of phospholipids in the solubilized opiate receptor complex. (B) Digitonin-NaCl was used for solubilization of receptors which were then labeled with [3H]diprenorphine. This complex could be chromatographed on hydroxylapatite, DEAE-sepharose and phenylboronate gels successively permitting an approximately 100-fold purification over the solubilized starting material.
D000818:Animals; D002047:Buprenorphine / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002540:Cerebral Cortex / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D004174:Diprenorphine / Q000378:metabolism; D010741:Phospholipases A; D054467:Phospholipases A2; D011957:Receptors, Opioid / Q000302:isolation & purification* / Q000378:metabolism; D012756:Sheep; D012995:Solubility; D010738:Type C Phospholipases
D011957:Receptors, Opioid; D004174:Diprenorphine; D002047:Buprenorphine; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases