title: Binding of Ca2+, Mg2+, and heparin by human serum amyloid P component in affinity capillary electrophoresis.
abstract: Human serum amyloid P component (SAP) is a glycoprotein circulating in the blood and found in association with all types of amyloid (malfolded potein aggregates) examined so far. Despite uncertainties regarding the precise function of SAP in vivo, the lectin-like properties of this Ca(2+)-activated protein with affinity for anionic saccharides and malfolded proteins are well known. The propensity to form homomeric penta- or decamers in solution and the selfaggregation in the presence of Ca(2+) as well as the tendency of SAP to attach to uncoated fused silica have precluded the analysis of SAP by microelectrophoretic methods. We now work out conditions to characterize the binding of Ca(2+) and Mg(2+) and the binding of heparin to SAP in the presence of divalent metal ions by ACE. The results show a strong binding of heparin (sub-muM apparent dissociation constants) even in the abscence of Ca(2+) at low ionic strength, pH 8.2. Also, a selective interaction with Ca(2+) compared with Mg(2+) is demonstrated. The approach will further the use of microelectrophoretic methods to examine the interactions of SAP with ligands of putative pathophysiological relevance such as lipopolysaccharides and misfolded proteins.
authors.affiliation: Department of Autoimmunology, Statens Serum Institut, Copenhagen S, Denmark. nhe@ssi.dk
authors.forename: Niels H H
authors.initials: NH
authors.lastname: Heegaard
authors.forename: Xinya
authors.initials: X
authors.lastname: He
authors.forename: Lars G
authors.initials: LG
authors.lastname: Blomberg
mesh_terms: D002118:Calcium / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D008274:Magnesium / Q000737:chemistry*; D000683:Serum Amyloid P-Component / Q000032:analysis* / Q000737:chemistry*
chemical_list: D000683:Serum Amyloid P-Component; D006493:Heparin; D008274:Magnesium; D002118:Calcium