Osmolytes improve the reconstitution of luteinizing hormone/human chorionic gonadotropin receptors into proteoliposomes.
Rat ovarian membrane luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor was reconstituted into proteoliposomes. The ability of sodium cholate to extract and reconstitute hCG binding activity was dependent on the protein/detergent ratio. Trypsinization of the LH/hCG receptor containing proteoliposomes indicated that approximately 57% of hCG binding sites were oriented extravesicularly. The presence of 20% glycerol or other osmolytes during reconstitution increased the accessibility of LH/hCG receptors but not the activity of adenylate cyclase in proteoliposomes. This beneficial effect was independent of any specific detergent or its presence during detergent solubilization of proteins. Dynamic properties of membranes were monitored by electron spin resonance of 16-, 12-, and 5-doxyl stearic acid. Reconstituted proteoliposomes contain less ordered membrane lipids than do native membranes. Addition of glycerol before reconstitution increased the order of lipid bilayer and shifted it to the physical state of the native membrane. These findings are consistent with the hypothesis that a rise in membrane ordering increases the accessibility of membrane-bound LH/hCG receptors.
Kolena
J
J
Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava.
Matejcíková
K
K
Srenkelová
G
G
D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D019826:Cholic Acid; D002793:Cholic Acids; D004578:Electron Spin Resonance Spectroscopy; D005990:Glycerol / Q000494:pharmacology*; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry*; D009994:Osmolar Concentration; D051381:Rats; D011919:Rats, Inbred Strains; D011967:Receptors, Gonadotropin / Q000378:metabolism*; D011974:Receptors, LH / Q000378:metabolism*; D012995:Solubility
D002793:Cholic Acids; D008051:Lipid Bilayers; D008081:Liposomes; D011967:Receptors, Gonadotropin; D011974:Receptors, LH; D000262:Adenylyl Cyclases; D019826:Cholic Acid; D005990:Glycerol