Identification of an autoantigen on the surface of apoptotic human T cells as a new protein interacting with inflammatory group IIA phospholipase A2.
One of the most studied secreted phospholipases A2 (sPLA2), the group IIA sPLA2, is found at high levels in inflammatory fluids of patients with autoimmune diseases. A characteristic of group IIA sPLA2 is its preference for negatively charged phospholipids, which become exposed on the extracellular leaflet of apoptotic cell membranes. We recently showed that low molecular weight heparan sulfate proteoglycans (HSPGs) and uncharacterized detergent-insoluble binding site(s) contribute to the enhanced binding of human group IIA PLA2 (hGIIA) to apoptotic human T cells. Using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry we now identify vimentin as the major HSPG-independent binding protein of hGIIA on apoptotic primary T lymphocytes. Vimentin is partially exposed on the surface of apoptotic T cells and binds hGIIA via its rod domain in a calcium-independent manner. Studies with hGIIA mutants showed that specific motifs in the interfacial binding surface are involved in the interaction with vimentin. The sPLA2 inhibitor LY311727, but not heparin, inhibited this interaction. In contrast, heparin but not LY311727 abrogated the binding of hGIIA to cellular HSPGs. Importantly, vimentin does not inhibit the catalytic activity of hGIIA. Altogether, the results show that vimentin, in conjunction with HSPGs, contributes to the enhanced binding of hGIIA to apoptotic T cells.
Boilard
Eric
E
Centre de Recherche en Rhumatologie et Immunologie, Université Laval, QC G1V 4G2, Canada.
Bourgoin
Sylvain G
SG
Bernatchez
Chantale
C
Surette
Marc E
ME
D020816:Amino Acid Motifs; D017209:Apoptosis; D001324:Autoantigens / Q000096:biosynthesis*; D001327:Autoimmune Diseases / Q000276:immunology; D001665:Binding Sites; D003902:Detergents / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D054501:Group II Phospholipases A2; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D009154:Mutation; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D014746:Vimentin / Q000737:chemistry / Q000502:physiology
D001324:Autoantigens; D003902:Detergents; D004791:Enzyme Inhibitors; D019812:Heparan Sulfate Proteoglycans; D007211:Indoles; C095553:LY 311727; D014746:Vimentin; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2