Kinetics of prothrombin-mediated binding of lupus anticoagulant antibodies to phosphatidylserine-containing phospholipid membranes: an ellipsometric study.
Antiphospholipid antibodies interact with phospholipid membranes via lipid binding plasma proteins, mostly, prothrombin and beta(2)-glycoprotein I. Using ellipsometry, we characterized prothrombin-mediated binding of lupus anticoagulant (LA) positive IgG, isolated from patients with antiphospholipid syndrome, to phosphatidylserine (PS)-containing membranes. LA IgG did not bind to membranes in the absence of prothrombin, but addition of prothrombin resulted in high-affinity binding of prothrombin-LA IgG complexes; half-maximal binding was attained at IgG and prothrombin concentrations of 10 microg/mL and 4 nM, respectively. Adsorption to membranes containing 10-40 mol % PS revealed that membrane-bound rather than solution-phase prothrombin determines the adsorption kinetics. Depletion of prothrombin and LA IgG from the solution results in rapid desorption which is strongly inhibited by addition of prothrombin but not of LA IgG. Prothrombin-mediated adsorption of monovalent Fab1 fragments prepared from patient LA IgG was negligible, indicating that monovalent interaction between prothrombin and LA IgG is weak. The kinetics of adsorption and desorption indicate that divalent binding of LA IgG to prothrombin at the lipid membrane occurs.
Willems
George M
GM
Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, The Netherlands. g.willems@carim.unimaas.nl
Janssen
Marie P
MP
Comfurius
Paul
P
Galli
Monica
M
Zwaal
Robert F A
RF
Bevers
Edouard M
EM
D017152:Antibodies, Antiphospholipid / Q000378:metabolism*; D001666:Binding Sites, Antibody; D015374:Biosensing Techniques; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G / Q000378:metabolism; D007164:Immunosorbents; D007700:Kinetics; D008051:Lipid Bilayers / Q000378:metabolism*; D016682:Lupus Coagulation Inhibitor / Q000276:immunology*; D010718:Phosphatidylserines / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D011516:Prothrombin / Q000737:chemistry* / Q000276:immunology / Q000502:physiology*; D012825:Silicon
D017152:Antibodies, Antiphospholipid; D007074:Immunoglobulin G; D007164:Immunosorbents; D008051:Lipid Bilayers; D016682:Lupus Coagulation Inhibitor; D010718:Phosphatidylserines; D011516:Prothrombin; D012825:Silicon