title: Capillary electrophoresis-based analysis of phospholipid and glycosaminoglycan binding by human beta2-glycoprotein I.
abstract: Human beta2-glycoprotein I (beta2gpI) is a phospholipid and heparin binding plasma glycoprotein involved in autoimmune diseases characterized by blood clotting disturbances (thrombosis) together with the occurrence of autoantibodies against beta2gpI. With the final goal of assessing autoantibody influence on binding interactions of beta2gpI we have studied the development of capillary electrophoresis (CE)-based assays for interactions of negatively charged ligands with beta2gpI. In the development of suitable conditions for analysis at neutral pH of this basic protein (pI about 8) we found the pH hysteresis behavior of fused silica surfaces useful since the protonated surface after an acid pre-wash counteracted protein adsorption efficiently in contrast to more laborious procedures including acrylamide/dimethylacrylamide coatings that did not permit analysis of this particular protein. This simple approach made estimates of heparin-beta2gpI interactions possible and the principle was shown also to work for detection of betagpI binding to anionic phospholipids. Utilizing the pH hysteresis effect may be a simple solution to the adsorption problems often encountered in analyses of proteins by CE.
authors.affiliation: Department of Chemistry, Karlstad University, SE-651 88 Karlstad, Sweden.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Bohlin
authors.forename: Ewa
authors.initials: E
authors.lastname: Kogutowska
authors.forename: Lars G
authors.initials: LG
authors.lastname: Blomberg
authors.forename: Niels H H
authors.initials: NH
authors.lastname: Heegaard
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding; D053482:beta 2-Glycoprotein I
chemical_list: D006023:Glycoproteins; D010743:Phospholipids; D053482:beta 2-Glycoprotein I