title: Effect of phospholipids on the reconstitution and thermal stability of delipidated rat ovarian luteinizing hormone/human chorionic gonadotropin receptors in proteoliposomes.
abstract: The role of lipids and a possible structure-functional alteration of delipidated LH/hCG receptor reconstituted into proteoliposomes was analyzed by thermal perturbation techniques. Delipidated receptor lost to a great extent its binding activity and thermal stability. The LH/hCG receptor was almost fully reactivated by the reconstitution into proteoliposomes with phosphatidylcholine (PC) and sphingomyelin (SpM) and partly with phosphatidylethanolamine (PE) and phosphatidic acid (PA). The heat inactivation profile of delipidated LH/hCG binding sites was shifted to a lower temperature by about 4 degrees C (T50 values). Thermal inactivation of the receptor by delipidation was entirely inverted by treatment with soybean PC, dioleoyl PC and dipalmitoyl PC and partially with SpM. The presence of negatively charged phospholipids, phosphatidylserine (PS), phosphatidylglycerol (PGl) and PA, did not change the heat-inactivation profile of the LH/hCG receptor modified the differential scanning calorimetric profile and the quenching of protein fluorescence characteristic for control proteoliposomes. Delipidation increased membrane lipid rigidity. Reconstitution of delipidated proteoliposomes with soybean PC, dioleoyl PC, PGl, PS and PA decreased, and that of dipalmitoyl PC, lysoPC, SpM and cholesterol increased the degree of fluorescence polarization of DPH of proteoliposomes. The different action of phospholipids on the reconstitution, thermal inactivation of the receptor and membrane lipid fluidity in proteoliposomes suggests that lipid fluidity is not related to the stabilizing action of phospholipids on the LH/hCG receptor.
authors.affiliation: Institute of Experimental Endocrinology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
authors.forename: J
authors.initials: J
authors.lastname: Kolena
authors.forename: S
authors.initials: S
authors.lastname: Scsuková
authors.forename: M
authors.initials: M
authors.lastname: Jezová
authors.forename: J
authors.initials: J
authors.lastname: Furdová
authors.forename: M
authors.initials: M
authors.lastname: Tatara
authors.forename: P
authors.initials: P
authors.lastname: Jasem
mesh_terms: D000818:Animals; D001665:Binding Sites; D002152:Calorimetry, Differential Scanning; D002462:Cell Membrane / Q000737:chemistry; D006063:Chorionic Gonadotropin / Q000378:metabolism; D004355:Drug Stability; D005260:Female; D005454:Fluorescence Polarization; D006358:Hot Temperature; D008081:Liposomes / Q000378:metabolism*; D008563:Membrane Lipids / Q000502:physiology; D010053:Ovary / Q000737:chemistry*; D010712:Phosphatidic Acids / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000494:pharmacology; D010714:Phosphatidylethanolamines / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D011974:Receptors, LH / Q000737:chemistry* / Q000502:physiology*; D013109:Sphingomyelins / Q000494:pharmacology; D013816:Thermodynamics
chemical_list: D006063:Chorionic Gonadotropin; D008081:Liposomes; D008563:Membrane Lipids; D010712:Phosphatidic Acids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D011974:Receptors, LH; D013109:Sphingomyelins