Chromatin-independent binding of serum amyloid P component to apoptotic cells.
Human serum amyloid P component (SAP) is a glycoprotein structurally belonging to the pentraxin family of proteins, which has a characteristic pentameric organization. Mice with a targeted deletion of the SAP gene develop antinuclear Abs, which was interpreted as evidence for a role of SAP in controlling the degradation of chromatin. However, in vitro SAP also can bind to phosphatidylethanolamine, a phospholipid which in normal cells is located mainly in the inner leaflet of the cell membrane, to be translocated to the outer leaflet of the cell membrane during a membrane flip-flop. We hypothesized that SAP, because of its specificity for phosphatidylethanolamine, may bind to apoptotic cells independent of its nuclear binding. Calcium-dependent binding of SAP to early, nonpermeable apoptotic Jurkat, SKW, and Raji cells was indeed observed. Experiments with flip-flopped erythrocytes confirmed that SAP bound to early apoptotic cells via exposed phosphatidylethanolamine. Binding of SAP was stronger to late, permeable apoptotic cells. Experiments with enucleated neutrophils, with DNase/RNase treatment of late apoptotic Jurkat cells, and competition experiments with histones suggested that binding of SAP to late apoptotic cells was largely independent of chromatin. Confocal laser microscopic studies indeed suggested that SAP bound to these apoptotic cells mainly via the blebs. Thus, this study shows that SAP binds to apoptotic cells already at an early stage, which raises the possibility that SAP is involved in dealing with apoptotic cells in vivo.
Familian
A
A
CLB, Sanquin Blood Supply Foundation and Laboratory for Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. A_Familian@clb.nl
Zwart
B
B
Huisman
H G
HG
Rensink
I
I
Roem
D
D
Hordijk
P L
PL
Aarden
L A
LA
Hack
C E
CE
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001667:Binding, Competitive; D002417:Cattle; D002843:Chromatin / Q000502:physiology*; D004492:Edetic Acid / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008024:Ligands; D009504:Neutrophils / Q000378:metabolism; D010949:Plasma / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D000683:Serum Amyloid P-Component / Q000302:isolation & purification / Q000378:metabolism*; D013997:Time Factors; D014407:Tumor Cells, Cultured
D002843:Chromatin; D008024:Ligands; D000683:Serum Amyloid P-Component; D004492:Edetic Acid