Structure-stabilizing effect of albumin on rat ovarian LH/hCG receptors.
The stabilizing effect of albumin on structure-functional alteration of LH/hCG receptors was analyzed by thermal perturbation technique. On exposing the membranes to bovine serum albumin (BSA) the heat inactivation profile of hCG-binding sites was shifted to a temperature higher by about 5 degreesC (T50 values). The receptor destabilizing action of arachidonic and oleic acids incorporated into ovarian membranes and reversal of this effect when BSA was used as fatty acid scavenger, may indicate that free fatty acids are responsible for the thermal instability of hCG-binding sites. This presumption was corroborated by digestion of membranes with phospholipase A2 (PLA2). This enzyme exerted effects on the thermal stability of the receptor protein resembling those observed upon insertion of fatty acids. The membrane fluidization induced by arachidonic acid can be reversed by BSA. However, alterations of lipid fluidity in membranes were not found to be a necessary prerequisite for stabilization of the LH/hCG receptor structure. Fluorescence quenching studies indicated that incorporation of oleic acid or digestion of membrane phospholipids with PLA2 elevated the accessibility of fluorophores for acrylamide. BSA scavenging of free fatty acids approached the quenching rate of control membranes. Analysis of fluorescence of membranes bound to monodansylcadaverine probe revealed that the negative surface charge derived from free fatty acids resulted in destabilization of the receptor protein. The effects of free fatty acids on membranes suggest that altered lipid-protein interactions may directly affect the stability of the LH/hCG receptor structure.
Kolena
J
J
Institute of Experimental Endocrinology, Slovak Academy of Sciences, Vlarska 3, 833 06, Bratislava, Slovak Republic. ueenkole@savba.sk
Jezová
M
M
Vranová
J
J
Scsuková
S
S
D000818:Animals; D016718:Arachidonic Acid / Q000737:chemistry; D001665:Binding Sites; D005260:Female; D006358:Hot Temperature; D008566:Membranes / Q000737:chemistry; D019301:Oleic Acid / Q000737:chemistry; D010053:Ovary / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D010741:Phospholipases A; D054467:Phospholipases A2; D051381:Rats; D017208:Rats, Wistar; D011974:Receptors, LH / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000494:pharmacology*
D011974:Receptors, LH; D012710:Serum Albumin, Bovine; D016718:Arachidonic Acid; D019301:Oleic Acid; D010741:Phospholipases A; D054467:Phospholipases A2