Anticoagulantly active heparin-like molecules from cultured fibroblasts.
Heparin-like molecules, isolated from cultured rat and human skin fibroblasts, accelerated the inactivation of purified human thrombin via purified human antithrombin. Only 15% of the biologic activity of the complex carbohydrates derived from human skin fibroblasts was expressed when the heparin-binding domain an antithrombin was chemically modified at the Trp 49 residue. Human skin fibroblasts were metabolically labeled with [35S]Na2SO4, and radiolabeled heparin-like molecules were isolated utilizing ion-exchange chromatography and fractionated into two separate populations employing immobilized antithrombin. The species which bound with high avidity to the affinity matrix represented about 12% of the radiolabeled heparin-like molecules, accounted for almost 95% of the initial anticoagulant activity, and exhibited a specific activity of 3.98 USP units/10(6) 35S-cpm.
D000991:Antithrombins / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism*; D006493:Heparin / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D019763:Heparin Lyase; D006801:Humans; D011133:Polysaccharide-Lyases / Q000378:metabolism; D012867:Skin; D013917:Thrombin / Q000378:metabolism
D000991:Antithrombins; D006493:Heparin; D013917:Thrombin; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase