Binding of human extracellular superoxide dismutase C to sulphated glycosaminoglycans.
The secretory enzyme extracellular superoxide dismutase (EC-SOD) occurs in at least three forms, which differ with regard to heparin affinity: A lacks affinity, B has intermediate affinity, and C has relatively strong affinity. The affinity of EC-SOD C for various sulphated glycosaminoglycans (GAGs) was assessed (a) by determining the concentration of NaCl required to release the enzyme from GAG-substituted Sepharose 4B and (b) by determining the relative potencies of the GAGs to release EC-SOD C from heparan sulphate-Sepharose 4B. Both methods indicated the same order of affinity. Heparin bound EC-SOD C about 10 times as avidly as the studied heparan sulphate preparation, which in turn was 10 and 150 times as efficient as dermatan sulphate and chondroitin sulphate respectively. Chondroitin sulphate showed weak interaction with EC-SOD C at physiological ionic strength. Heparin subfractions with high or low affinity for antithrombin III were equally efficient. The binding of EC-SOD C to heparin-Sepharose was essentially independent of pH in the range 6.5-9; below pH 6.5 the affinity increased, and beyond pH 9.5 there was a precipitous fall in affinity. The inhibitory effect of NaCl on the binding of EC-SOD C to GAGs indicates that the interaction is of electrostatic nature. EC-SOD C carries a negative net charge at neutral pH, and it is suggested that the binding occurs between the negative charges of the GAG sulphate groups and a structure in the C-terminal end of the enzyme that has a cluster of positive charges. These results are compatible with the notion that heparan sulphate proteoglycans on cell surfaces or in the intercellular matrix may serve to bind EC-SOD C in tissues.
Karlsson
K
K
Department of Clinical Chemistry, Umeå University Hospital, Sweden.
Lindahl
U
U
Marklund
S L
SL
D000345:Affinity Labels; D005110:Extracellular Space / Q000201:enzymology; D006025:Glycosaminoglycans / Q000378:metabolism*; D006493:Heparin; D006497:Heparan Sulfate; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D012685:Sepharose / Q000031:analogs & derivatives; D012965:Sodium Chloride; D013482:Superoxide Dismutase / Q000378:metabolism*
D000345:Affinity Labels; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D012685:Sepharose; D012965:Sodium Chloride; D013482:Superoxide Dismutase; C037405:heparin-sepharose