Microsomal opiate receptors differ from synaptic membrane receptors in proteolytic sensitivity.
We have found that opiate receptors in smooth microsomal fractions differ from synaptic membrane-associated receptors in proteolytic sensitivity. With 3 proteases of different substrate specificities (trypsin, chymotrypsin and S. griseus protease) smooth microsomal opiate receptors from rat brain were consistently less sensitive to limited proteolysis than were synaptic membrane receptors. Thiamine pyrophosphatase, a luminal Golgi membrane marker enzyme, exhibited a similar resistance to S. griseus protease in microsomal preparations, while microsomal Na+/K+-ATPase (ouabain-sensitive) was readily destroyed by trypsin. We also discovered that smooth microsomal opiate receptors co-migrate with both Golgi membrane and endoplasmic reticulum marker proteins on equilibrium density gradients under isopycnic conditions. Electron microscopic examination of the Golgi-enriched fraction showed the typical cisternae frequently associated with isolated Golgi membranes. Synaptic junctions, presynaptic membranes, myelin and mitochondria were conspicuously absent from this fraction. Since the microsomes isolated in vitro showed similar topography to those in vivo, the binding sites for opiates could be localized on the luminal surface membranes of the microsomal fractions. The exquisite sensitivity of synaptic membrane opiate receptors to proteolysis suggests that these receptors are found on the extracellular surface of the synaptic junction.
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002421:Caudate Nucleus / Q000378:metabolism; D002458:Cell Fractionation; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D008861:Microsomes / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011957:Receptors, Opioid / Q000187:drug effects / Q000302:isolation & purification*; D013379:Substrate Specificity; D013570:Synaptic Membranes / Q000378:metabolism* / Q000648:ultrastructure; D013788:Thalamus / Q000378:metabolism
D011957:Receptors, Opioid; D010447:Peptide Hydrolases