title: Proteolytic modification of the heparin-binding affinity of extracellular superoxide dismutase.
abstract: The heparin-binding affinity of the tetrameric extracellular superoxide dismutase (EC-SOD) is a result of the cooperative effect of the heparin-binding domains of the subunits, located in the hydrophilic, strongly positively charged C-terminal ends. EC-SOD C, the high-heparin-affinity type, exposed to immobilized trypsin and plasmin was found to rapidly lose its affinity for heparin, without any loss of enzymic activity or major change in molecular mass as judged by size-exclusion chromatography. Heparin and dextran sulphate 5000 inhibited the proteolysis, suggesting that EC-SOD C sequestered by heparan sulphate proteoglycan in vivo is partially protected against proteolysis. The loss of heparin-affinity occurred with the stepwise formation of intermediates, and the pattern upon chromatography on heparin-Sepharose and subsequent immunoblotting was compatible with the notion that the changes are due to sequential truncations of heparin-binding domains from subunits composing the EC-SOD tetramers. A similar pattern with intermediates and apparent truncations has previously been found with EC-SOD of human plasma. The findings show that the unique design of the heparin-binding domain of EC-SOD allows easy modification of the heparin-affinity by means of limited proteolysis, and suggest that such proteolysis is a major contributor to the heterogeneity in heparin-affinity of EC-SOD in mammalian plasma.
authors.affiliation: Department of Clinical Chemistry, Umeå University Hospital, Sweden.
authors.forename: K
authors.initials: K
authors.lastname: Karlsson
authors.forename: A
authors.initials: A
authors.lastname: Edlund
authors.forename: J
authors.initials: J
authors.lastname: Sandström
authors.forename: S L
authors.initials: SL
authors.lastname: Marklund
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D013379:Substrate Specificity; D013482:Superoxide Dismutase / Q000378:metabolism*
chemical_list: D006493:Heparin; D013482:Superoxide Dismutase