RSS-Feed abonnieren
DOI: 10.1055/s-0038-1661594
A Computer Assisted Method to Obtain the Prothrombin Activation Velocity in Whole Plasma Independent of Thrombin Decay Processes
Publikationsverlauf
Received 24. Februar 1988
Accepted 02. Mai 1986
Publikationsdatum:
13. Juli 2018 (online)
Summary
A method is described that, on the basis of the time course of amidolytic activity after the triggering of thrombin generation in normal plasma, allows the calculation of the velocity of prothrombin conversion independent of thrombin inactivating processes.
It is shown how the reaction constants for the α2M-dependent and the α2M-independent thrombin inactivation processes can be obtained in a sample of whole plasma.
The method is verified by demonstrating that the experimentally observed time courses of residual prothrombin and of α2M-thrombin complex coincide with those calculated from the time course of amidolytic activity, and by showing that the course of prothrombin conversion in plasma without α2-macroglobulin or AT III is adequately described if the α2M or AT Ill-dependent breakdown constants are taken zero in the calculations.
It appears that the inactivation of thrombin, endogenously generated in whole plasma, is about half as fast as that of exogenous thrombin added to the plasma.
A computer program is presented that carries out the relevant calculations.
-
References
- 1 Wagner RH, Graham JB, Penick GD, Brinkhous KM. Estimation of prothrombin by the two stage method. In: Blood coagulation, Hemorrhage and Thrombosis Tocantins ML, Kazal LA. (eds) Academic Press; New York - London: 1965
- 2 Biggs R, Macfarlane RG. Human blood coagulation and its disorders. Blackwell Scientific Publications; Oxford: 1967
- 3 Hemker HC, Hemker PW, vd Torren K, Devilee PP, Hermens WTh, Loeliger EA. The evaluation of the two-stage prothrombin assay. Thromb Diathes Haemorrh 1971; 25: 545-554
- 4 Pekelharing CA. Over de betrekking van het fibrineferment van het bloedserum tot de nucleoproteide van het bloedplasma. Versl Kon Acad Wetensch Amsterdam 1895; 3: 272-297
- 5 Travis J, Salvesen GS. Human plasma proteinase inhibitors. Ann Rev Biochem 1983; 52: 655-709
- 6 Rosenberg RD, Harpel PC. α2-Macroglobulin and antithrombinheparin cofactor: modulators of hemostatic and inflammatory reactions. Prog Hemost Thromb 1976; 3: 145-198
- 7 Barret AJ, Starkey PM. The interaction of α2macroglobulin with proteinase. Biochem J 1973; 133: 709-715
- 8 Harpel PC. Studies on human plasma α2-macroglobulin enzyme interactions. Evidence for proteolytic modification of the subunit chain structure. J Exp Med 1973; 138: 508-521
- 9 Rinderknecht H, Feling RM, Geokas MC. Effect of α2-macroglobulin in some kinetic parameters of trypsin. Biochim Biophys Acta 1975; 377: 150-165
- 10 Fischer AM, Tapon-Bretaudière J, Bros A, Josso F. Respective roles of antithrombin III and α2-macroglobulin in thrombin inactivation. Thromb Haemostas 1981; 45: 51-54
- 11 Josso F, Prou-Wartelle O. Exploration de l’hémostase. In: Techniques en hématologie Alagille D. et al. (eds) pp 101-208 Flamarion; Paris: 1972
- 12 van Dam-Mieras MC E, Muller AD, van Dieijen G, Hemker HC. Methods of enzymatic analysis Enzymes 3: Peptidases, proteinase and their inhibitors. Verlag Chemie, Weinheim 1984; 5: 352-394
- 13 Ødegard OR, Lie M. On the use of chromogenic substrates for the study of coagulation inhibitors. Haemostasis 1978; 7: 121-126
- 14 Owren PA, Aas K. The control of dicumarol therapy and the quantitative determination of prothrombin and proconvertin. Scand J Clin Lab Invest 1951; 3: 201-218
- 15 Hendrix H, Lindhout T, Mertens K, Engels W, Hemker HC. Activation of human prothrombin by stoichiometric levels of staphy-locoagulase. J Biol Chem 1983; 258: 3637-3644
- 16 Steinbuch M, Audran R. Isolement de l’immunoglobuline IgG du plasma humain à l’aide de l’acide caprylique. Rev Franç Etud clin biol 1969; 14: 1054-1058
- 17 Lindhout T, Baruch D, Schoen P, Franssen J, Hemker HC. Thrombin generation and inactivation of antïthrombin III and heparin. (Submitted for publication)
- 18 Kawabata S, Morita T, Iwanaga S, Igarashi H. Staphylocoagulase-binding region in human prothrombin. J Biochem 1985; 97: 329-331
- 19 Rosenberg RD, Damus PS. The purification and mechanism action of human antithrombin-heparin cofactor. J Biol Chem 1973; 248: 6490-6505
- 20 Downing MR, Bloom JW B, Mann KG. Comparison of the inhibition of thrombin by three plasma protease inhibitors. Biochemistry 1978; 17: 2649-2653
- 21 Rosing J, Zwaal RF A, Tans G. Formation of meizothrombin as an intermediate in Factor Xa-catalyzed prothrombin activation. J Biol Chem 1986 (In press)
- 22 Esmon CT, Jackson CM. The conversion of prothrombin to thrombin. IV. The function of the fragment 2 region during activation in the presence of factor V. J Biol Chem 1974; 249: 7791-7797
- 23 Jesty J. The kinetics of inhibition of thrombin by antithrombin in the presence of components of the hemostatic systems. Blood 1985; 66: 1189-1195
- 24 Hemker HC, Willems G, Béguin S. Thrombinoscope, a personal computer program for the analysis of thrombin generation curves. Computers in Biol and Med. (Submitted)