Abstract
Thrombokinase has been isolated from bovine plasma by a procedure which begins with the highly purified product of a previously described method, chromatographs it on DEAE-cellulose, and then fractionates it by continuous flow electrophoresis, yielding 0.2 mg per liter of oxalated plasma. The electrophoretic fraction has shown a single boundary in the ultracentrifuge; and its esterase activity on toluenesulfonylarginine methyl ester has been about the same as that of thrombokinase previously isolated by repeated electrophoretic fractionations. Thrombokinase is a euglobulin with minimum solubility near pH 5.0. It is most stable within the pH range 7.5 to 9.5; but there is also a peak in the stability curve near pH 1.8. A few micrograms of thrombokinase per milliliter can activate prothrombin in the presence of EDTA. A few thousandths of a microgram causes rapid production of thrombin in the system: prothrombin, thrombokinase, calcium chloride, phosphatide, "accelerator." But, thrombokinase has less than 1/175 the proteolytic activity of crystallized trypsin.
Full Text
The Full Text of this article is available as a PDF (796.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ALEXANDER B. Coagulation, hemorrhage and thrombosis. N Engl J Med. 1955 Mar 24;252(12):484–494. doi: 10.1056/NEJM195503242521205. [DOI] [PubMed] [Google Scholar]
- CHRISTENSEN L. R., SMITH D. H., Jr Plasminogen purification by acid extraction. Proc Soc Exp Biol Med. 1950 Aug;74(4):840–844. doi: 10.3181/00379727-74-18063. [DOI] [PubMed] [Google Scholar]
- CONNOR W. E., CARTER J. R. Influence of soybean phosphatide on blood coagulation and its use in the thromboplastin generation test. Proc Soc Exp Biol Med. 1958 Jan;97(1):38–43. doi: 10.3181/00379727-97-23640. [DOI] [PubMed] [Google Scholar]
- HADLEY G. G., LARSON N. L. Use of sequestrene as an anticoagulant. Am J Clin Pathol. 1953 Jun;23(6):613–618. [PubMed] [Google Scholar]
- HYUN B. H., DAWSON E. A., BUTCHER J., CUSTER R. P. Studies on soybean phosphatide (inosithin) as a platelet substitute. Stability and effective concentrations in the thromboplastin generation test. Am J Clin Pathol. 1960 Mar;33:209–213. doi: 10.1093/ajcp/33.3.209. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- MILSTONE J. H. Fractionation of plasma globulin for prothrombin, thrombokinase, and accessory thromboplastin. J Gen Physiol. 1951 Sep;35(1):67–87. doi: 10.1085/jgp.35.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MILSTONE J. H. Preparation of thrombokinase from bovine plasma. J Gen Physiol. 1959 Mar 20;42(4):665–676. doi: 10.1085/jgp.42.4.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MILSTONE J. H. Separation of thrombin from thrombokinase by continuous flow paper electrophoresis. Proc Soc Exp Biol Med. 1959 Aug-Sep;101:660–662. doi: 10.3181/00379727-101-25052. [DOI] [PubMed] [Google Scholar]
- MILSTONE J. H. TAMe esterase activity of blood thrombokinase after repeated electrophoretic fractionations. Proc Soc Exp Biol Med. 1960 Feb;103:361–363. doi: 10.3181/00379727-103-25519. [DOI] [PubMed] [Google Scholar]
- NEWLANDS M. J., WILD F. Sources of platelet factor for the thromboplastin generation test. Nature. 1955 Nov 5;176(4488):885–886. doi: 10.1038/176885a0. [DOI] [PubMed] [Google Scholar]
- OWEN C. A., Jr, THOMPSON J. H., Jr Soybean phosphatides in prothrombin-consumption and thromboplastin-generation tests: their use in recognizing "thrombasthenic hemophilia". Am J Clin Pathol. 1960 Mar;33:197–208. doi: 10.1093/ajcp/33.3.197. [DOI] [PubMed] [Google Scholar]
- SPAET T. H., KROPATKIN M. Effect of intravenous soy bean phosphatides on blood coagulation in rabbits. Proc Soc Exp Biol Med. 1957 Jul;95(3):492–495. doi: 10.3181/00379727-95-23263. [DOI] [PubMed] [Google Scholar]