Binding of human extracellular-superoxide dismutase C to cultured cell lines and to blood cells.
The high heparin-affinity subtype C of the secretory enzyme extracellular-superoxide dismutase (EC-SOD) was found to bind to cultured mammalian cells, forming an equilibrium between the cells and the medium. To anchorage-dependent cell lines, binding apparently occurred both to the glycocalyx of the cell surfaces and to the sub- and intercellular matrix produced by the cells. Heparan sulfate proteoglycan appeared to be the principal binding substance. The binding capacities of anchorage-dependent cultures were very high, and at maximal binding the amount of EC-SOD C activity associated with the exterior of the cells was several-fold higher than the endogenous intracellular SOD activity. Half-maximal binding occurred at about 8 micrograms/ml EC-SOD C. At low, nonsaturating, physiologic EC-SOD C concentrations, the enzyme concentration in the glycocalyx of cells may be several thousand times higher than in the medium. All 14 investigated anchorage-dependent cell lines, including endothelial cells, bound EC-SOD C avidly. The 10 suspension-growing cell lines were all weaker binders. Blood monomorphonuclear leukocytes and platelets bound little EC-SOD C, whereas no significant binding to neutrophil leukocytes, to erythrocytes and to E. coli could be demonstrated. The findings are compatible with the notion that EC-SOD C in the vasculature forms an equilibrium between plasma and heparan sulfate in the glycocalyx of the endothelium. Furthermore, tissue EC-SOD is probably distributed between heparan sulfate on the surface of most cell types in the organs and in the interstitial matrix. The binding pattern suggests that EC-SOD C has the potential to protect most normal cells in the body and the interstitial matrix, without protecting microorganisms lacking affinity, and without interfering with superoxide radicals produced at the surface of activated neutrophil leukocytes.
Karlsson
K
K
Department of Clinical Chemistry, Umeå University Hospital, Sweden.
Marklund
S L
SL
D000818:Animals; D001773:Blood Cells / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000378:metabolism; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism*; D013696:Temperature; D014407:Tumor Cells, Cultured
D011508:Chondroitin Sulfate Proteoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D013482:Superoxide Dismutase