title: Lipoprotein lipase induces catabolism of normal triglyceride-rich lipoproteins via the low density lipoprotein receptor-related protein/alpha 2-macroglobulin receptor in vitro. A process facilitated by cell-surface proteoglycans.
abstract: Bovine milk lipoprotein lipase (LPL) induced binding, uptake, and degradation of 125I-labeled normal human triglyceride-rich lipoproteins by cultured mutant fibroblasts lacking LDL receptors. The induction was dose-dependent and occurred whether LPL and 125I-lipoproteins were added to incubation media simultaneously or LPL was allowed to bind to cell surfaces, and unbound LPL was removed by washing prior to the assay. Lipolytic modification of lipoproteins did not appear to be necessary for increased catabolism because the effect of LPL was not prevented by inhibitors of LPL's enzymatic activity, p-nitrophenyl N-dodecylcarbamate or phenylmethylsulfonyl fluoride. However, the effect was abolished by boiling LPL prior to the assay suggesting that major structural features of LPL were required. Also, LPL-induced binding to cells was blocked by an anti-LPL monoclonal antibody but not by antibodies that are known to block apolipoprotein E- or B-100-mediated binding to low density lipoprotein (LDL) receptors. This indicates that LPL itself mediated 125I-lipoprotein binding to cells. Cellular degradation of 125I-lipoproteins was partially or completely blocked by two previously described ligands for the LDL receptor-related protein/alpha 2-macroglobulin receptor (LRP): activated alpha 2-macroglobulin (alpha 2M*), and the 39-kDa receptor-associated protein. These data implicated LRP as mediating LPL-induced lipoprotein degradation and were confirmed by showing that LPL's effects were prevented by an immunoaffinity-isolated polyclonal antibody against LRP. Furthermore, LPL promoted binding of 125I-lipoproteins to highly purified LRP in a solid-phase assay. Heparin or heparinase treatment of cells markedly decreased LPL-induced binding, uptake, and degradation of lipoproteins, but had no effect on catabolism of alpha 2M*. Thus, cell-surface proteoglycans were obligatory participants in the effects of LPL but were not required for LRP-mediated catabolism of alpha 2M*. Taken together, these in vitro findings establish that through interaction with cell-surface proteoglycans, LPL induces catabolism of normal human triglyceride-rich lipoproteins via LRP.
authors.affiliation: Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242.
authors.forename: D A
authors.initials: DA
authors.lastname: Chappell
authors.forename: G L
authors.initials: GL
authors.lastname: Fry
authors.forename: M A
authors.initials: MA
authors.lastname: Waknitz
authors.forename: L E
authors.initials: LE
authors.lastname: Muhonen
authors.forename: M W
authors.initials: MW
authors.lastname: Pladet
authors.forename: P H
authors.initials: PH
authors.lastname: Iverius
authors.forename: D K
authors.initials: DK
authors.lastname: Strickland
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000378:metabolism; D001057:Apolipoproteins E / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D019763:Heparin Lyase; D006801:Humans; D007700:Kinetics; D008071:Lipoprotein Lipase / Q000378:metabolism*; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008892:Milk / Q000201:enzymology; D010920:Placenta / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000494:pharmacology; D011247:Pregnancy; D011509:Proteoglycans / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000378:metabolism*; D011973:Receptors, LDL / Q000235:genetics / Q000378:metabolism; D012867:Skin / Q000378:metabolism*; D014280:Triglycerides / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D001057:Apolipoproteins E; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011509:Proteoglycans; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D014280:Triglycerides; D006493:Heparin; D008071:Lipoprotein Lipase; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase