Autoantigen Ro52 directly interacts with human IgG heavy chain in vivo in mammalian cells.
Previously, when we used in vivo yeast two-hybrid and in vitro protein-protein interaction analyses, we demonstrated a direct interaction between autoantigen Ro52 and the human IgG heavy chain. This interaction occurred in the absence of antibody-antigen specific interaction. Here, by employing a novel strategy, we further demonstrated that Ro52 co-localized with IgG in transfected mammalian cells. The co-localization was specific to IgG1 but not IgG3. Co-immunoprecipitating IgG with Ro52 from transfected cell lysates suggested that protein complex containing Ro52 and IgG contributed to the in vivo co-localization. In addition, IgG from normal human serum was shown to bind to the surface of apoptotic keratinocytes and the binding could be competitively blocked by 50-fold excesses of IgG1, not IgG3. With a direct binding study, we also demonstrated that IgG1 could bind to the surface of apoptotic cells while IgG3 bound barely. This binding was not competed by Fcgamma fragments indicating a non-Fcgamma receptor mediated interaction. Finally, in a competition analysis the addition of GST-RFP could reduce the IgG binding to the cell surface. Thus, we suggested that the binding of IgG to the apoptotic keratinocytes might be mediated through the interactions with the surface exposed Ro52. The potential role of forming this protein complex on the apoptotic cells will be discussed.
Yang
Y S
YS
Department of Internal Medicine, Pulmonary and Critical Care Division, UT Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9034, USA. yih-sheng.yang@utsouthwestern.edu
Yang
M C
MC
Wang
B
B
Weissler
J C
JC
D000818:Animals; D017209:Apoptosis; D001324:Autoantigens / Q000276:immunology*; D002460:Cell Line; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D015603:Keratinocytes / Q000502:physiology; D008322:Mammals; D051379:Mice; D011485:Protein Binding; D020733:RNA, Small Cytoplasmic; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins / Q000276:immunology*; D020798:Two-Hybrid System Techniques; D000099207:SS-A Antigen
D001324:Autoantigens; D007143:Immunoglobulin Heavy Chains; D020733:RNA, Small Cytoplasmic; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D000099207:SS-A Antigen; C116006:RO60 protein, human