RSS-Feed abonnieren
DOI: 10.1055/s-0038-1654857
Purification and Kinetic Studies on a Circulating Anticoagulant in a Suspected Case of Lupus Erythematosus*
This investigation was supported (in part) by a Public Health Service Research Career Program Award (# AM-8661) from the National Institute of Arthritis and Metabolic Diseases and by a State of Washington Initiative 171 Funds for Research in Biology and Medicine.Publikationsverlauf
Publikationsdatum:
24. Juli 2018 (online)

Summary
1. A circulating anticoagulant in a suspected case of lupus erythematosus has been highly purified by a combination of Sephadex gel filtration and DEAE cellulose chromatography.
2. The inhibitor is a gamma globulin with a sedimentation coefficient of 6.6 Svedberg units.
3. For its anticoagulant action, the lupus inhibitor requires a co-factor which is present both in the lupus and normal blood.
4. The cofactor is located in the gamma globulins fraction which is relatively heat stable, but less than the inhibitor, at 56° C.
5. The active lupus inhibitor (inhibitor + cofactor) is not species specific against human prothrombin.
6. Working with highly purified systems, the active inhibitor is found to inhibit prothrombin conversion by formed prothrombin activator. It does not appear to inhibit the formation of prothrombin activator nor does it affect purified prothrombin.
** Present address: Department of Medicine, The Jewish Hospital, St. Louis, Missouri (U.S.A.).
-
References
- 1 Bachmann F, Duckert F, Geiger M, Baer P, Koller F. Differentation of the factor VII complex. Studies on the Stuartprower factor. Thrombos. Diathes. haemorrh. (Stuttg) 01: 169 1957;
- 2 Bachmann F, Duckert F, Koller F. The Stuart-Prower factor Assay and its clinical significance. Thrombos. Diathes. haemorrh. (Stuttg) 02: 24 1958;
- 3 Biggs R, Denson KWE. The mode of action of a coagulation inhibitor in the blood of two patients with disseminated lupus erythematosus (DLE). : Brit. J. Haemat. 10: 198 1964;
- 4 Biggs R, Macfarlane R. G. Human blood coagulation and its disorders. 3rd ed.,. 378 F.A. Davis Co; Philadelphia: 1962
- 5 Biggs R, Macfarlane R. G. Human blood coagulation and its disorders. 3rd ed.,. 391 F.A. Davis Co; Philadelphia: 1962
- 6 Biggs R, Macfarlane R. G. Human blood coagulation and its disorders. 3rd. ed.,. 385 F.A. Davis Co; Philadelphia: 1962
- 7 Brecher G, Conkite E. P. Morphology and enumeration of human blood platelets. Amer. J. clin. Path. 23: 15 1953;
- 8 Breckenridge R. T, Ratnoff O. D. Studies on the site of action of a circulating anticoagulant in disseminated lupus erythematosus. Amer. J. Med. 35: 813 Dec. 1963;
- 9 Glauss A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta haemat. (Basel) 187: 373 1956;
- 10 Vermylen C, Verstraete M. Antithrombin V: Critical evaluation of its assessment and properties. Thrombos. Diathes. haemorrh. (Stuttg) 05: 267 1960;
- 11 Duckert F, Jung E, Schmerling D. H. Hitherto undescribed congenital haemorrhagic diathesis probably due to fibrin stabilizing factor deficiency. Thrombos. Diathes. haemorrh. (Stuttg) 05: 179 1960;
- 12 Duckert F, Yin E. T, Straub W. Separation and purification of the blood clotting factors by means of chromatography and electrophoresis. Preparative methods. VIII Colloquium. Protides of biological fluids. Sint-Janshospitaal, Brugge, Belgium. May 1960
- 13 Fahey J. L, McCoy P. F, Goulian M. Chromatography of serum proteins in normal and pathological sera: The distribution of protein-bound carbohydrate and cholesterol, siderophilin, thyroxinbinding protein, B12-binding protein, alkaline and acid phosphatases, radio- iodinated albumin and myeloma proteins. J. clin. Invest 37: 272 1958;
- 14 Flodin P, Killander J. Fractionation of human-serum proteins by gel filtration. Biochim. biophys. Acta (Amst) 63: 403 1962;
- 15 Frick P. G. Acquired circulating anticoagulants in systemic “Collagen disease”. Autoimmune thromboplastin deficiency. Blood 10: 691 1955;
- 16 Gaston L. W, Evanger A. E. The requirement for calcium in the thromboplastin generation test. J. Lab, clin. Med. 64: 501 1964;
- 17 Gaston L. W, Mach B. F, Beck W. S. Hemophilia A and concurrent factor VII deficiency. New Engl. J. Med. 264: 1078 1961;
- 18 Gornall A. G, Bardawill G. J, David M. N. Determination of serum proteins by means of the biuret reaction. J. Biol. Chem. 177: 751 1949;
- 19 Grabar P. Immunoelectrophoresis analysis. Methods of biochemical analysis. Glick D. ed New York: Interscience Publishers Inc; 7 1 1959
- 20 Hanahan D. J, Dittmer J. C, Warashina E. A column chromatographic separation of classes of phospholipides. J. Biol. Chem. 228: 685 1957;
- 21 Hjort P. F. Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scand. J. clin. Lab. Invest 09 (Supp. 27): 1 1957;
- 22 Hougie C. Naturally occuring species specific inhibitor of human prothrombin in lupus erythematosus. Proc. Soc. exp. Biol. Med. 116: 359 1964;
- 23 Kappeler R. Das Verhalten von Faktor V im Serum unter normalen und pathologischen Bedingungen. Z. klin. Med. 153: 103 1955;
- 24 Koller F, Loeliger A, Duckert F. Experiments on a new clotting factor. Factor VII. Acta haemat. (Basel) 06: 1 1951;
- 25 Langdeil R. D, Wagner R. H, Brinkhous K. M. Effect of antihemophilic factor on One- stage clotting tests. A presumptive test for hemophiliacs and a simple one-stage antihemophilic factor assay procedure. J. Lab. clin. Med. 41: 637 1953;
- 26 Loeliger A. Prothrombin as a co-factor of the circulating anticoagulant in systemic lupus erythematosus ? Thrombos. Diathes. haemorrh. (Stuttg) 03: 237 1959;
- 27 Margolius Jr. A, Jackson DPO, Ratnoff D. Circulating anticoagulants: Study of 40 cases and review of literature. Medicine 40: 145 1961;
- 28 Conley C. L, Hartmann R. C. A haemorrhagic disorder caused by circulating anticoagulant in patients with disseminated lupus erythematosus. J. clin. Invest 31: 621 1952;
- 29 Papahadjopoulos D, Yin E. T, Hanahan D. J. Purification and properties of bovine factor X: Molecular changes during activation. Biochem. 03: 1931 1964;
- 30 Papahadjopoulos D, Yin E. T, Hanahan D. J. To be published..
- 31 Proctor R. R, Rapaport S. I. The partial thromboplastin time with kaolin. Amer. J. clin. Path. 36: 212 1961;
- 32 Rapaport S. I, Ames S. B, Duvall B. J. A plasma coagulation defect in systemic lupus erythematosus arising from hypoprothrombinemia combined with antiprothrombinase activity. Blood 15: 212 1960;
- 33 Ratnoff O. D, Menzies C. A New method for the determination of fibrinogen in small samples of plasma. J. Lab. clin. Med. 37: 316 1951;
- 34 Rouser G, Baumann A. J, KitchevsJcy G, Heller D, O’Brien J. S. Quantitative chromato- graphysic fractionation of complex lipid mixture: Brain lipids. J. Am. Oil Chem. Soc. 38: 544 1961;
- 35 Schachman H. K. Ultracentrifugation in biochemistry. 55 Academic Press; New York and London: 1959
- 36 Shulman N. R, Hearon J. Z. Kinetics of conversion of prothrombin to thrombin by biological activators. I. Demonstration of a prothrombin derivative which reacts with proteolytic enzyme inhibitors. J. Biol. Chem. 238 Jan. 1963
- 37 Streuli F. On the purification and conversion of human prothrombin. Thrombos. Diathes. haemorrh. (Stuttg) 03: 194 1959;
- 38 United States Dept, of Health, Education and Welfare. Public Health Service. Laboratory procedures for modern syphilis serology. 1 1961
- 39 Waaler B. A. Contact activation in the intrinsic blood clotting system. Scand. J. clin. Lab. Invest 11 (supp. 37): 01: 1959
- 40 Wintrobe M. M. Clinical hematology. 5th ed.,. 419 Lea and Febiger; Philadelphia: 1961
- 41 WillTcens R. F, Decher J. L. Rheumatoid arthritis with serologic evidence suggesting lupus erythematosus: Clinical, serologic and chromatographic studies. Arth. Rheumat. 06: 720 1963;
- 42 Yin E. T. Kinetics of human blood coagulation induced by trypsin. Thrombos. Diathes. haemorrh. (Stuttg) 12: 307 1964;