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DOI: 10.1055/s-0038-1657240
Activation of Factor IX by Factor XIa - A Spectrophotometric Assay for Factor IX in Human Plasma[*]
Publication History
Received 26 April 1982
Accepted 30 June 1982
Publication Date:
13 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/198202/lookinside/thumbnails/10-1055-s-0038-1657240-1.jpg)
Summary
The activation of Factor IX by partially purified Factor XIa was followed by active site titration, gelelectrophoresis and by a spectrophotometric assay. The assay is based on the finding that the rate of Factor X activation in the presence of phospholipid and Ca2+ is linear in time and proportional to the amount of Factor IXa present and can be determined with the chromogenic substrate S2222. Conditions were found that allowed complete activation of Factor IX in human plasma by Factor XIa. The amount of Factor IXa present in the plasma sample can be determined with the spectrophotometric assay and is proportional with the amount of plasma present. In plasma from patients receiving vitamin-K antagonists reduced Factor IX activity is found with the spectrophotometric assay, and the new assay method may be useful in monitoring oral anticoagulant therapy.
Keywords
Active site titration of F IXa - Activation of F IX by F XIa - Spectrophotometric assay for F IXa - Chromogenic substrate - Factor XIa - Factor IX - Hemophilia B* Part of this work was presented by G.T. to the Rijksuniversiteit Limburg in partial fulfilment of the requirements for the Ph. D. degree. The chromogenic substrates used in this work were in part donated by KABI AB, Stockholm, Sweden. Part of this work was presented at the VIIIth International Congress on Thrombosis and Haemostasis, Toronto, Canada, July 1981.
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References
- 1 Van Dieijen G, Tans G, Rosing J, Hemker HC. The role of phospholipid and factor VIIIa in the activation of bovine factor X. J Biol Chem 1981; 256: 3433-3442
- 2 Davie EW, Fujikawa K. Basic mechanisms in blood coagulation. Ann Rev Biochem 1975; 44: 799-828
- 3 Jackson CM, Nemerson Y. Blood coagulation. Ann Rev Biochem 1980; 49: 765-811
- 4 Fujikawa K, Thompson AR, Legaz ME, Meyer RG, Davie EW. Isolation and characterization of bovine factor IX (Christmas factor). Biochemistry 1973; 12: 4938-4945
- 5 Østerud B, Flengsrud R. Purification and some characteristics of the coagulation factor IX from human plasma. Biochem J 1975; 145: 469-474
- 6 Andersson CO, Borg H, Miller-Andersson M. Purification and characterization of human factor IX. Thromb Res 1975; 7: 451-459
- 7 Rosenberg JS, Beeler DL, Rosenberg RD. Activation of human prothrombin by highly purified human factors V and Xa in the presence of human antithrombin. J Biol Chem 1975; 250: 1607-1617
- 8 Suomela H. Human coagulation factor IX. Isolation and characterization. Eur J Biochem 1976; 71: 145-154
- 9 Di Scipio RG, Kurachi K, Davie EW. Activation of human factor IX (Christmas factor). J Clin Invest 1978; 1528-1538
- 10 Katayama K, Ericsson LH, Enfield DL, Walsh KA, Neurath H, Davie EW, Titani K. Comparison of amino acid sequence of bovine coagulation factor IX (Christmas factor) with that of other vitamin Independent plasma proteins. Proc Natl Acad Sci USA 1979; 76: 4990-4994
- 11 Fujikawa K, Legaz ME, Kato H, Davie EW. The mechanism of activation of bovine factor IX (Christmas factor) by bovine factor XIa (activated plasma thromboplastin antecedent). Biochemistry 1974; 13: 4508-4516
- 12 Lindquist PA, Fujikawa K, Davie EW. Activation of bovine factor IX (Christmas factor) by factor XIa (activated plasma thromboplastin antecedent) and a protease from Russell’s viper venom. J Biol Chem 1978; 253: 1902-1909
- 13 Østerud B, Rapaport SI. Activation of factor IX by the reaction product of tissue factor and factor VII: additional pathway for initiating blood coagulation. Proc Natl Acad Sci USA 1974; 12: 5260-5264
- 14 Zur M, Shastri K, Nemerson Y. Kinetics of the tissue factor pathway of coagulation: Activation of factor IX. Blood 52 (Suppl. 01) 1978; 198
- 15 Byrne R, Amphlett GW, Castellino FJ. Metal ion specificity of the conversion of bovine factors IX, IXα, and IXaα to bovine factor IXaβ . J Biol Chem 1980; 255: 1430-1435
- 16 Veltkamp JJ, Drion EF, Loeliger EA. Detection of the carrier state in hereditary coagulation disorders I. Thrombos Diathes Haemorrh 1968; 19: 279-308
- 17 Byrne R, Castellino FJ. The influence of metal ions in the activation of bovine factor IX by the coagulation protein of Russell’s viper venom. Arch Biochem Biophys 1978; 190: 687-692
- 18 Silverberg SA, Nemerson Y, Zur M. Kinetics of the activation of bovine coagulation factor X by components of the extrinsic pathway: Kinetic behavior of two-chain factor VII in the presence and absence of tissue factor. J Biol Chem 1977; 252: 8481-8488
- 19 Byrne R, Link RP, Castellino FJ. A kinetic evaluation of activated bovine blood coagulation factor IX toward synthetic substrates. J Biol Chem 1980; 255: 5336-5341
- 20 Cuatrecasas P. Protein purification by affinity chromatography; derivatizations of agarose and polyacrylamide beads. J Biol Chem 1970; 245: 3059-3065
- 21 Fujikawa K, Legaz ME, Davie EW. Bovine factor X1 and X2 (Stuart factor). Isolation and characterization. Biochemistry 1972; 11: 4882-4891
- 22 Nossel HL. The contact phase of blood coagulation. Blackwell scientific Publications; Oxford: 1964
- 23 Schiffman S, Theodor I, Rapaport SI. Separation from Russell’s viper venom of one fraction reacting with factor X and another reacting with factor V. Biochemistry 1969; 8: 1397-1405
- 24 Fujikawa K, Legaz ME, Davie EW. Bovine factor X (Stuart factor). Mechanism of activation by a protein from Russell’s viper venom. Biochemistry 1972; 11: 4892-4899
- 25 Sanders H. Preparative isolation of phosphatidylserine from brain. Biochim Biophys Acta 1967; 144: 485-487
- 26 de Kruijff B, Cullis PR, Radda GK. Differential scanning calorimetry and 31P NMR studies on sonicated and unsonicated phosphatidylcholine liposomes. Biochim Biophys Acta 1975; 406: 6-20
- 27 Bender ML, Begué-Canton ML, Blakeley RL, Brubacher LJ, Feder J, Gunter CR, Kézdy FJ, Killheffer JV, Thomas HM, Miller CG, Roeske RW, Stoops JK. The determination of the concentration of hydrolytic enzyme solutions: α-chymotrypsin, trypsin, papain, elastase, subtilisin and acetyl Cholinesterase. J Am Chem Soc 1966; 88: 5890-5913
- 28 Bender ML, Kézdy FJ, Wedler FC. Chymotrypsin. Enzyme concentration and kinetics. J Chem Educ 1967; 44: 84-88
- 29 Chase Jr T, Shaw E. Comparison of the esterase activities of trypsin, plasmin and thrombin on guanidinobenzoate ester. Titration of the enzymes. Biochemistry 1969; 8: 2212-2224
- 30 Smith RL. Titration of activated bovine factor X. J Biol Chem 1973; 218: 2418-2423
- 31 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
- 32 Fairbanks G, Stech TL, Wallach DFH. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 1971; 10: 2606-2617
- 33 Østerud B, Bouma BN, Griffin JH. Human blood coagulation factor IX. Purification properties and mechanism of activation by activated factor XI. J Biol Chem 1978; 253: 5946-5951
- 34 Suttie JW. Mechanism of action of vitamin K synthesis of γ-carboxy glutamic acid. Crit Rev Biochem 1980; 8: 191-223
- 35 Suttie JW, Jackson CM. Prothrombin Structure, Activation and Biosynthesis. Physiol Rev 1977; 57: 1-70
- 36 Lindhout MJ, Kop-Klaassen BHM, Hemker HC. The effect of γ-carboxyglumate residues on the enzymatic properties of the activated bloodclotting factor X. Biochim Biophys Acta 1978; 533: 342-354