title: The alpha -chains of C4b-binding protein mediate complex formation with low density lipoprotein receptor-related protein.
abstract: C4b-binding protein (C4BP) is a heparin-binding protein that participates in both the complement and hemostatic system. We investigated the interaction between C4BP and low density lipoprotein receptor-related protein (LRP), an endocytic receptor involved in the catabolism of various heparin-binding proteins. Both plasma-derived C4BP and recombinant C4BP consisting of only its alpha-chains (rC4BPalpha) bound efficiently to immobilized LRP, as determined by surface plasmon resonance analysis. Complementary, two distinct fragments of LRP, i.e. clusters II and IV, both associated to immobilized rC4BPalpha, and binding could be inhibited by the LRP antagonist receptor-associated protein. Further analysis showed that association of rC4BPalpha to LRP was inhibited by heparin or by anti-C4BP antibody RU-3B9, which recognizes the heparin-binding region of the C4BP alpha-chains. In cellular degradation experiments, LRP-expressing fibroblasts effectively degraded (125)I-labeled rC4BPalpha, whereas their LRP-deficient counterparts displayed a 4-fold diminished capacity of degrading (125)I-rC4BPalpha. Finally, initial clearance of C4BP in mice was significantly delayed upon co-injection with receptor-associated protein. In conclusion, our data demonstrate that the alpha-chains of C4BP comprise a binding site for LRP. We propose that LRP mediates at least in part the catabolism of C4BP and, as such, may regulate C4BP participation in complement and hemostatic processes.
authors.affiliation: Laboratory for Thrombosis and Haemostasis, Department of Haematology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: Westein
authors.forename: Cecile V
authors.initials: CV
authors.lastname: Denis
authors.forename: Bonno N
authors.initials: BN
authors.lastname: Bouma
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Lenting
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D003169:Complement Inactivator Proteins; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006023:Glycoproteins; D006493:Heparin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D011485:Protein Binding; D011951:Receptors, Complement / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D020349:Surface Plasmon Resonance; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011951:Receptors, Complement; D011994:Recombinant Proteins; D012965:Sodium Chloride; D006493:Heparin; D005982:Glutathione Transferase