Binding, degradation, and utilization of plasma high density and low density lipoproteins for progesterone production in cultured rat luteal cells.
These studies were intended to examine the binding and degradation of plasma lipoprotein fractions and the utilization of lipoprotein-bound cholesterol for progesterone production in cultured rat luteal cells. These cells bound [125I]human low density lipoprotein ([125I]iodo-hLDL),[125I]human high density lipoprotein ([125I]iodo-hHDL), and [125I]iodorat HDL ([125I] iodo-rHDL) with high affinity. The equilibrium dissociation constants of the binding of labeled rHDL, hHDL, and hLDL were 90.5, 78.3, and 36.8 micrograms/ml, respectively. All three lipoproteins were also degraded in a concentration-dependent manner, with apparent Km values of 18.3, 17.5, and 22.4 micrograms/ml for rHDL, hHDL, and hLDL, respectively. The degradation of the lipoproteins was inhibited by lysosomotropic agents, transglutaminase inhibitors, and metabolic inhibitors, suggesting that these lipoproteins undergo internalization and lysosomal degradation. In addition, all three lipoproteins also augmented the hCG-stimulated steroidogenesis. When cells were incubated with reconstituted LDL (the cholesterol ester in the LDL was replaced with [3H]cholesteryl linoleate) and the steroids produced identified, incorporation of tritium label in the progesterone fraction was observed in a time- and concentration-dependent manner. The incorporation of tritium into progesterone was increased by hCG and inhibited by an excess of unlabeled LDL in the incubation medium. These results show that the ovarian cells use lipoproteins as a source of cholesterol for steroidogenesis through receptor-mediated uptake and internalization, and the evidence suggests that the lipoproteins are intracellularly degraded.
D000818:Animals; D002352:Carrier Proteins; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D003338:Corpus Luteum / Q000378:metabolism*; D005260:Female; D007700:Kinetics; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism*; D011374:Progesterone / Q000096:biosynthesis*; D011555:Pseudopregnancy / Q000378:metabolism; D016601:RNA-Binding Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000378:metabolism; D011973:Receptors, LDL; D018110:Receptors, Lipoprotein
D002352:Carrier Proteins; D006063:Chorionic Gonadotropin; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D016601:RNA-Binding Proteins; D011956:Receptors, Cell Surface; D011973:Receptors, LDL; D018110:Receptors, Lipoprotein; C031616:high density lipoprotein receptors; C075197:high density lipoprotein binding protein; D011374:Progesterone