Binding of complement proteins C1q and C4bp to serum amyloid P component (SAP) in solid contra liquid phase.
Serum amyloid P component (SAP), a member of the conserved pentraxin family of plasma proteins, binds calcium dependently to its ligands. The authors investigated SAPs interaction with the complement proteins C4b binding protein (C4bp) and C1q by ELISA, immunoelectrophoresis and electron microscopy. Binding of these proteins to SAP was demonstrated when SAP was immobilized using F(ab')2 anti-SAP, but not when SAP reacted with these proteins in liquid phase; thus the binding to human SAP was markedly phase state dependent. Presaturation of solid phase SAP with heparin, which binds SAP with high affinity, did not interfere with the subsequent binding of C4bp or C1q to SAP. In contrast, collagen I and IV showed partial competition with the binding of C1q to SAP. Using fresh serum, immobilized native SAP bound C4bp whereas binding of C1q/C1 could not be demonstrated. Altogether the results indicate that firm binding of C1q and C4bp to SAP requires that SAP is presented on a solid phase, that C1q and C4bp react with sites distinct from the heparin binding site, and that C1q and collagen I share binding sites on SAP. Immobilized native SAP, aggregated SAP and SAP-heparansulphate complexes induced no detectable complement activation.
Sørensen
I J
IJ
Department of Medical Microbiology, Odense University, Denmark.
Nielsen
E H
EH
Andersen
O
O
Danielsen
B
B
Svehag
S E
SE
D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism*; D003167:Complement Activation; D015922:Complement C1q / Q000378:metabolism*; D004586:Electrophoresis / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D016167:Integrin alphaXbeta2; D011485:Protein Binding; D011817:Rabbits; D000683:Serum Amyloid P-Component / Q000378:metabolism*
D002352:Carrier Proteins; D015922:Complement C1q; D006497:Heparan Sulfate; D016167:Integrin alphaXbeta2; D000683:Serum Amyloid P-Component