Delipidation of a beta-adrenergic receptor preparation and reconstitution by specific lipids.
The role of lipids in the function of membrane receptors for hormones and neurotransmitters is still obscure. To gain information on this subject, a delipidated receptor preparation was developed. The beta-adrenergic receptor from turkey erythrocyte membranes was solubilized in deoxycholate and was freed extensively of phospholipids and of cholesterol by gel filtration. The delipidated preparation, after removal of the detergent, showed little, if any, ligand binding to the receptor as measured with the beta-adrenergic antagonist [125I] iodocyanopindolol. Readdition of soybean lipids restored specific radioligand binding. The lipid reconstituted receptor demonstrated agonist and antagonist binding affinities which were not very different from those of the native receptor. The receptor also retained its ability to function, as demonstrated by transfer to a foreign adenylate cyclase system. The delipidated receptor preparation lent itself conveniently to study the requirement for specific lipids in restoration of agonist and antagonist binding. Phosphatidylethanolamine restored maximal binding. Acidic phospholipids and sphingomyelin were inefficient in reconstitution of the receptor. The effect of cholesterol addition was also investigated. Binding was dramatically increased when a cholesterol ester was added in mixture with the acidic phospholipids, cardiolipin or phosphatidylinositol. Further studies unexpectedly revealed that reconstitution of the delipidated receptor is not exclusively dependent on the addition of a phospholipid; a mixture of 1-monooleylglycerol with cholesteryl hemisuccinate restored binding as efficiently as phosphatidylethanolamine. The presently described preparation should be useful in elucidating the part played by lipids in the action of the receptor in the adenylate cyclase system.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D003840:Deoxycholic Acid; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D007942:Leukemia, Experimental / Q000378:metabolism; D008563:Membrane Lipids / Q000097:blood / Q000302:isolation & purification*; D051379:Mice; D010743:Phospholipids / Q000097:blood / Q000302:isolation & purification*; D011941:Receptors, Adrenergic / Q000302:isolation & purification*; D011943:Receptors, Adrenergic, beta / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility; D014422:Turkeys
D008563:Membrane Lipids; D010743:Phospholipids; D011941:Receptors, Adrenergic; D011943:Receptors, Adrenergic, beta; D003840:Deoxycholic Acid; D000262:Adenylyl Cyclases