Expression and function of the low density lipoprotein receptor-related protein (LRP) in mammalian central neurons.
The low density lipoprotein receptor-related protein (LRP) is a multifunctional cell surface receptor, expressed in liver, that binds with high affinity and endocytoses several structurally and functionally distinct ligands, including apolipoprotein E-activated beta-migrating very low density lipoprotein, tissue-type plasminogen activator, and alpha 2-macroglobulin. Here using in situ hybridization and quantitative RNase protection assays, we show that LRP is also expressed throughout the brain. LRP message is particularly high in the cerebellum, cortex, hippocampus, and brain stem. In addition, we demonstrate that a 39-kDa protein which copurifies with LRP and regulates the binding of other ligands to LRP is also expressed throughout the brain. Interestingly, expression of the 39-kDa message is approximately 100-fold of that found in liver, suggesting that the activity of LRP is more tightly regulated in brain tissue than in liver. Using primary cultures of isolated postnatal cortical neurons, [35S]methionine biosynthetic labeling and immunoprecipitation, we also demonstrate the de novo biosynthesis of LRP, the 39-kDa protein, as well as tissue-type plasminogen activator. Finally, using radioligand binding as well as fluorescent ligand binding and uptake studies, we show that LRP is functional in cortical neurons. These results taken together thus demonstrate expression of functional LRP in neuronal cells and suggest a potential role for LRP in brain protein and lipoprotein metabolism, development, and regeneration.
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Edward Mallinckrodt Department of Pediatrics and Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110.
Maksymovitch
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EA
Nerbonne
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JM
Schwartz
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D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D015870:Gene Expression; D017403:In Situ Hybridization; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D000511:alpha-Macroglobulins / Q000378:metabolism
D017931:DNA Primers; D018076:DNA, Complementary; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D013462:Sulfur Radioisotopes; D000511:alpha-Macroglobulins; D008715:Methionine