RSS-Feed abonnieren
DOI: 10.1055/s-0038-1654095
The Conversion of Prethrombin to Thrombin[*]
Publikationsverlauf
Publikationsdatum:
26. Juni 2018 (online)

Summary
Purified prethrombin was converted to thrombin in an activating system consisting of purified autoprothrombin C, purified Ac-globulin, lipid and calcium chloride. With the concentration of calcium and pH fixed, the effect of varying the other 3 pro-coagulants was studied.
Conditions influencing the conversion of prethrombin to thrombin were determined. Autoprothrombin C is the enzyme, but with it alone activation was very slow. Best results were obtained with the simultaneous presence of autoprothrombin C, Ac-globulin and lipid. Reducing any one of the three toward zero concentration decreased the rate and yield of thrombin generation. In association with rapid thrombin generation Ac-globulin and autoprothrombin C were represented in approximately a 1:1 molar ratio. In that combination of Ac-globulin and autoprothrombin C the Michaelis constant for autoprothrombin C with prethrombin as a substrate was 3.14 × 10-6M.
* This investigation was supported by a research grant HE-03424-09 from the National Heart Institute, National Institutes of Health, IT. S. Public Health Service.
-
References
- 1 Seegers W. H, McGlaughry R. I, Fahey J. L. Some properties of purified prothrombin and its activation with sodium citrate. Blood 5: 421 1950;
- 2 Lorand L, Alkjaersig N, Seegers W. H. Carbohydrate and nitrogen distribution during the activation of purified prothrombin in sodium citrate solution. Arch. Biochem. Biophys 45: 312 1953;
- 3 Marciniak E, Seegers W. H. Experiments with prothrombin, prethrombin, autoprothrombin III, and plasmas with prothrombin related deficiencies. New Instanbul Contr. clin. Sci 8: 117 1965;
- 4 Marciniak E, Seegers W. H. Prethrombin as a new sub-unit of prothrombin. Nature (Lond.) 209: 621 1966;
- 5 Seegers W. H, Aoki N, Marciniak E. Autoprothrombin C and the generation of thrombin from prothrombin activation mixtures. New Instanbul Contr. clin. Sci 5: 170 1962;
- 6 Seegers W. H, Gole E. R, Aoki N, Harmison G. R. Separation of autoprothrombin III from bovine prothrombin preparations. Canad. J. Biochem 42: 229 1964;
- 7 Seegers W. H, Cole E. R, Harmison C. R, Marciniak E. Purification and some properties of autoprothrombin C. Canad. J. Biochem 41: 1047 1963;
- 8 Aoki N, Harmison C. R, Seegers W. H. Properties of bovine Ac-globulin concentrates and methods of preparation. Canad. J. Biochem 41: 2409 1963;
- 9 Cole E. R, Marciniak E, Seegers W. H. Procedures for the quantitative determination of autoprothrombin C. Thrombos. Diathes. haemorrh. (Stuttg.) 8: 434 1962;
- 10 Hecht E. R, Cho M. H, Seegers W. H. Thromboplastin: Nomenclature and preparation of protein-free material different from platelet factor 3 or lipid activator. Amer. J. Physiol 105: 584 1958;
- 11 Ware A. G, Seegers W. H. Plasma Accelerator Globulin: Partial purification, quantitative determination and properties. J. biol. Chem 172: 699 1948;
- 12 Ware A. G, Seegers W. H. Two-stage procedure for the quantitative determination of prothrombin concentration. Amer. J. clin. Path 19: 471 1949;
- 13 Dixon M. The determination of enzyme inhibitor constants. Biochem. J 55: 170 1953;
- 14 Lineweaver H, Burk D. The determination of enzyme dissociation constants. J. Amer, chem. Soc 56: 658 1934;
- 15 Seegers W. H, Heene D. L, Marciniak E. Activation of purified prothrombin in ammonium sulfate solutions: Purification of autoprothrombin C. Thrombos. Diathes. haemorrh. (Stuttg.) 15: 1 1966;
- 16 Mertz E. T, Seegers W. H, Smith H. P. Prothrombin, thromboplastin and thrombin: Quantitative interrelationships. Proc. Soc. exp. Biol. (N. Y.) 42: 604 1939;
- 17 Loomis E. G, Seegers W. H. Purified prothrombin: Factors which influence its activation. Arch. Biochem 5: 265 1944;
- 18 Seegers W. H, Marciniak E. Some activation characteristics of the prethrombin sub-unit of prothrombin. Life Sciences 4: 1721 1965;
- 19 Ware A. G, Seegers W. H. Serum Ac-globulin: formation from plasma Ac-globulin; role in blood coagulation; partial purification; properties; and quantitative determination. Amer. J. Physiol 152: 567 1948;
- 20 Seegers W. H, Cole E. R, Aoki N. Function of Ac-globulin and lipid in blood clotting. Canad. J. Biochem 41: 2441 1963;
- 21 Seegers W. H, Kagami M. The separation of autoprothrombin Ic from bovine prothrombin preparations. Canad. J. Biochem 42: 1249 1964;
- 22 Seegers W. H, Alkjaersig N, Johnson S. A. Formation of autoprothrombin in solutions containing purified prothrombin and purified platelet factor 3. Amer. J. Physiol 181: 589 1955;
- 23 Mammen E. F, Thomas W. R, Seegers W. H. Activation of purified prothrombin to autoprothrombin I or autoprothrombin II (platelet cofactor II) or autoprothrombin II-A. Thrombos. Diathes. haemorrh. (Stuttg.) 5: 218 1960;
- 24 Seegers W. H, Johnson S. A. Conversion of prothrombin to autoprothrombin II (platelet cofactor II) and its relation to the blood clotting mechanisms. Amer. J. Physiol 184: 259 1956;