title: Interaction of lipoprotein lipase and receptor-associated protein.
abstract: Receptor-associated protein (RAP) is a recognized chaperone/escort protein for members of the low density lipoprotein receptor family. In this report, we show that RAP binds to lipoprotein lipase (LPL) and may play a role in the maturation of LPL. Binding of highly purified RAP to LPL was demonstrated in vitro by solid phase assays, surface plasmon resonance, and rate zonal centrifugation. The dissociation constant for this interaction measured by the first two techniques ranged between 2.4 and 13 nM, values similar to those reported for the binding of RAP to LRP or gp330. The specificity of the interaction was demonstrated by competition with a panel of LPL monoclonal antibodies. Rate zonal centrifugation demonstrated the presence of a stable complex with an apparent Mr consistent with the formation of a complex between monomeric LPL and RAP. RAP x LPL complexes were co-immunoprecipitated in adipocyte lysates or from solutions of purified LPL and RAP. The interaction was also demonstrated in whole cells by cross-linking experiments. RAP-deficient adipocytes secreted LPL with a specific activity 2.5-fold lower than the lipase secreted by control cells. Heparin addition to cultured RAP-deficient adipocytes failed to stimulate LPL secretion in the medium, suggesting defective binding of the lipase to the plasma membrane. These studies demonstrate that RAP binds to LPL with high affinity both in purified systems and cell extracts and that RAP-deficient adipocytes secrete poorly assembled LPL. A function of RAP may be to prevent premature interaction of LPL with binding partners in the secretory pathway, namely LRP and heparan sulfate proteoglycan.
authors.affiliation: Division of Nutritional Sciences, Cornell University, Ithaca, New York 14853, USA.
authors.forename: Shallee
authors.initials: S
authors.lastname: Page
authors.forename: Andrea
authors.initials: A
authors.lastname: Judson
authors.forename: Kristan
authors.initials: K
authors.lastname: Melford
authors.forename: André
authors.initials: A
authors.lastname: Bensadoun
mesh_terms: D017667:Adipocytes / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry; D007700:Kinetics; D026563:LDL-Receptor Related Protein-Associated Protein / Q000737:chemistry* / Q000378:metabolism; D008071:Lipoprotein Lipase / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D011485:Protein Binding; D051381:Rats; D020349:Surface Plasmon Resonance
chemical_list: D003432:Cross-Linking Reagents; D019812:Heparan Sulfate Proteoglycans; D026563:LDL-Receptor Related Protein-Associated Protein; D008071:Lipoprotein Lipase