[Activation by heparin-Sepharose of prothrombin conversion to prethrombin 1].
Using affinity chromatography on heparin-Sepharose, homogeneous prothrombin containing no factor Xa or thrombin was separated into three components differing in their affinities for the bioadsorbent. The major component 2 eluted with 0.35 M NaCl was found to contain prothrombin (Mr 80000) and prethrombin 1 (Mr 60000). Component 1 not bound by heparin contained fragment 1 of prothrombin (Mr 25 000), whereas component 3 with a higher affinity for the bioadsorbent contained factor IX (Mr 52 000). Rechromatography of component 2 provided further evidence for prothrombin modification to prethrombin 1 by heparin-Sepharose. Blocking of endogenous thrombin of prothrombin by diisopropylfluorophosphate did not affect the modification. Heparin-Sepharose probably induced changes in prothrombin conformation and the formation of a catalytic center responsible for prothrombin splitting to prethrombin 1. Heparin-Sepharose can be used for separation of prothrombin proteolytic products by thrombin and for isolation of prethrombin 1 and prothrombin fragment 1.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology*; D008970:Molecular Weight; D011516:Prothrombin / Q000302:isolation & purification / Q000378:metabolism*; D012685:Sepharose
D004792:Enzyme Precursors; D011516:Prothrombin; D006493:Heparin; D012685:Sepharose; C012007:prethrombins
Aktivatsiia geparin-sefarozoĭ prevrashcheniia protrombina v pretrombin 1.