Inhibition of activated protein C by aprotinin and the use of the insolubilized inhibitor for its purification.
The study of the interaction between activated protein C (APC) and non-plasmatic inhibitors allowed us to demonstrate that aprotinin is a potent competitive inhibitor of APC with a Ki of 1.35 mumol/L. It was possible to adsorb immunopurified protein C (PC) activated by venom activator to insolubilized aprotinin and to recover the active enzyme after elution by HCl 0.1 N or by a chaotropic ion, for example KSCN 3 mol/L. The interaction involved the active-site of the enzyme since PC and DIP-APC did not bind to the matrix. Thus, APC could be purified, after activation, in a one-stage procedure out of a mixture of protein such as a prothrombin complex concentrate.
Centre National de Transfusion Sanguine, Les Ulis, France.
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Taby
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Chabbat
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Steinbuch
D000577:Amides / Q000378:metabolism; D007611:Aprotinin / Q000378:metabolism / Q000494:pharmacology*; D001780:Blood Coagulation Tests; D002846:Chromatography, Affinity; D006801:Humans; D011480:Protease Inhibitors / Q000494:pharmacology; D011486:Protein C / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D012995:Solubility
D000577:Amides; D011480:Protease Inhibitors; D011486:Protein C; D007611:Aprotinin