Rapid sulfopropyl-disk chromatographic purification of bovine and human thrombin.
Thrombin, from either a crude commercial preparation (bovine) or a prothrombin activation mixture (human), was purified by sulfopropyl-disk chromatography to homogeneity in a rapid and convenient single-step procedure. The yield of both proteinases was greater than 85%. Purified bovine and human thrombin had sp act of 2500 and 3000 "NIH" units/mg, respectively. Human thrombin was more reactive than bovine thrombin with these active site-directed probes: phenylmethylsulfonyl fluoride, dansyl-Glu-Gly-Arg-chloromethylketone, and human heparin cofactor II. The sulfopropyl-disk chromatographic method is a useful and rapid technique to prepare milligram quantities of highly purified bovine and human thrombin.
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D000990:Antithrombin III / Q000494:pharmacology; D002417:Cattle; D002845:Chromatography / Q000379:methods*; D003619:Dansyl Compounds; D006023:Glycoproteins / Q000494:pharmacology; D015844:Heparin Cofactor II; D006801:Humans; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors / Q000302:isolation & purification*
D000590:Amino Acid Chloromethyl Ketones; D003619:Dansyl Compounds; D006023:Glycoproteins; D010664:Phenylmethylsulfonyl Fluoride; C030824:dansylglutamyl-glycyl-arginine chloromethyl ketone; D015844:Heparin Cofactor II; D000990:Antithrombin III; D013917:Thrombin