Oxidation and biotinylation of beta 2 glycoprotein I glycan chains induce an increase in its affinity for anionic phospholipids similar to that obtained by the addition of anti-beta 2 glycoprotein I or anti-cardiolipin antibodies.
Binding of beta 2 glycoprotein I (beta2GPI) to apoptotic cells plays a key role in the opsonization of apoptotic bodies and the formation of antiphospholipids antibodies. Here, we describe the binding of beta2GPI to apoptotic cells using beta2GPI labelled with biotin-hydrazide (beta2GPI-bh) after oxidation of its glycan chains. Flow cytometry analyses and confocal microscopy showed that beta2GPI-bh, contrary to native beta(GPI, bound to apoptotic cells, either permeable or non-permeable to propidium iodide (PI), as did annexin-V-FITC. But, in the absence of divalent ions, beta2GPI-bh, contrary to annexin V, was still able to bind to apoptotic cells. Binding equilibrium studies, performed on solid-state anionic phospholipids (AnPL), revealed that beta2GPI-bh had a greater apparent affinity for AnPL than native beta2GPI. In presence of the anti-beta2GPI mAb 8C3, the ability of native beta2GPI to bind to AnPL was increased and binding to apoptotic PI+ and PI- CEM cells was observed whereas binding of beta2GPI-bh was barely affected by the addition of 8C3. However, the 8C3-enhanced ability of native beta2GPI to bind to AnPL was still weaker than that of beta2GPI-bh. It is not clear why the oxidation and biotinylation of glycan chains of beta2GPI increases its affinity for AnPL, but it seems that if such oxidative process occurs naturally, it could participate in enhancing antiphospholipid formation.
d'Angeac
Arnaud Dupuy
AD
Laboratoire de Chimie Biomoléculaire, UMR CNRS 5032, UM II, ENSCM, 8 rue de l'Ecole Normale 34296 Montpellier Cedex 5, France. dangeac@univ-montp2.fr
Stefas
Ilias
I
Duperray
Christophe
C
Rucheton
Marcel
M
Graafland
Hubert
H
Montero
Jean-Louis
JL
Chicheportiche
Robert
R
D000818:Animals; D017153:Antibodies, Anticardiolipin / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D017209:Apoptosis; D001665:Binding Sites; D019921:Biotinylation; D002460:Cell Line; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D010084:Oxidation-Reduction; D010743:Phospholipids / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D053482:beta 2-Glycoprotein I
D017153:Antibodies, Anticardiolipin; D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D010743:Phospholipids; D011134:Polysaccharides; D011994:Recombinant Proteins; D053482:beta 2-Glycoprotein I