Purification and properties of guinea pig antithrombin III.
Guinea pig antithrombin III has been purified from plasma by sequential heparin-Sepharose affinity chromatography, DE-52 cellulose chromatography, isoelectric focussing, and Sephadex G-100 gel filtration chromatography. The final product was homogeneous as judged by sodium dodecyl sulfate disc gel electrophoresis. Purification was 202-fold with a yield of 41%. Antiproteinase activity of antithrombin III was determined by progressive inactivation of thrombin coagulant and amidolytic activity. Heparin cofactor activity was demonstrated by immediate inactivation of thrombin by antithrombin III in the presence of minute quantities of heparin. It also could be demonstrated that thrombin inactivation by antithrombin III occurs by formation of a bimolecular complex whose rate of formation is markedly enhanced by minute quantities of heparin.
D000818:Animals; D000990:Antithrombin III / Q000302:isolation & purification* / Q000494:pharmacology; D001780:Blood Coagulation Tests; D002846:Chromatography, Affinity; D006168:Guinea Pigs / Q000097:blood*; D006493:Heparin / Q000494:pharmacology; D008970:Molecular Weight; D011480:Protease Inhibitors; D013917:Thrombin / Q000037:antagonists & inhibitors
D011480:Protease Inhibitors; D000990:Antithrombin III; D006493:Heparin; D013917:Thrombin