Subscribe to RSS
DOI: 10.1055/s-0038-1653568
On the Functional Resynthesis of Thromboplastin (“TP”) from its Fractions and Various Substitutes
Publication History
Publication Date:
10 June 2018 (online)
Summary
Recombination of the low active lipid fraction with the inactive non-lipid fraction of brain thromboplastin has been proved to be just as successful in restoring the original high thromboplastic activity as the recombination of the corresponding material obtained from lung thromboplastin.
The properties of the individual components, of thromboplastin, its fractions obtained by different ways, and of their substitutes were studied comparatively; the influence of the solvents used was also investigated.
Experiences obtained were collected to prepare a high active and stable brain thromboplastin.
Only the lipid- and non-lipid fraction obtained by pyridin is successful in regenerating the original thromboplastic activity, in contrast with the reunion of both fractions produced by Sephadex G-25, Sephadex LH-20 and butanol.
The lipid fraction can be fully replaced by the lipid activator, PE from soy-beans and partly by synthetic dioleoyl PE, however this is not possible with natural and synthetic PC and lyso PC.
The results of experiments to replace the non-lipid fraction by various amino acids, peptides or proteins were poor.
A full regeneration of the thromboplastic activity with the suspended fractions is only successful after a previous solution of the components in pyridin or chloroform and evaporation.
The rebuilt of the original thromboplastic activity was complete only as long as contaminated non-lipid fractions were used. Non-lipid fractions, quantitatively free of lipids, were ineffective.
1) Present address: State University of Utrecht, Department of Biochemistry, Utrecht, The Netherlands.
-
References
- 1 Bell W. N, Alton H. G. A brain extract as a substitute for platelet suspensions in the thromboplastin generation test. Nature (Lond) 174: 880 1954;
- 2 Bordet J, Delange L. Sur la nature du cytozyme. Ann. Instit. Pasteur 27: 341 1913;
- 3 Chargaff E, Moore D. H, Bendich A. Ultracentrifugal isolation from lung tissue of a macro - molecular protein component with thromboplastic properties. J. biol. Chem 145: 593 1942;
- 4 Deutsch E, Irsigler K, Lomoschitz H. Studien über Gewebsthromboplastin. 1. Reinigung, Chemische Charakterisierung und Trennung in einen Eiweiß- und Lipoidanteil. Thrombos. Diathes. haemorrh. (Stuttg) 12: 12 1964;
- 5 Hecht E. Zur Kenntnis der Blutgerinnung, 1. Mitt. Zur Methodik der Bestimmung der Blutgerinnungszeit. Acta med. scand 102: 79 1939;
- 6 Hecht E. Zur Kenntnis der Blutgerinnung, 2. Mitt. Versuche zur quantitativen Bestimmungsmethodik der Blutgerinnungszeit. Acta med. scand 102: 85 1939;
- 7 Hecht E. Untersuchungen über die chemische Natur der Gerinnungsfaktoren. Sang 21: 486 1950;
- 8 Hecht E. Prothrombinebepalingen en thrombokinasen. Ned. Tijdschr. Geneesk 97: 1996 1953;
- 9 Hecht E. Eine systematische Untersuchung über die Bedeutung der Lipoide für die Blutgerinnung, 2. Mitt. Der Einfluß der Diaminophosphatide und deren Spaltungsprodukt Sphingosin. Acta haemat. (Basel) 09: 237 1953;
- 10 Hecht E. Zur Kenntnis der Thrombokinasen. Biochem. Z 326: 325 1955;
- 11 Hecht E. Studien über den lipoiden Aktivator der Blutgerinnung. Thrombos Diathes. haemorrh. (Stuttg) 01: 380 1957;
- 12 Hecht E. Lipids in Blood Clotting. Charles C. Thomas; Springfield, Ill., U.S.A: 1965
- 13 Hecht E. Chemical nature of human brain thromboplastin. Nature (Lond) 214: 197 1967;
- 14 Hecht E, Oosterbaan-van Lit W. L. Über die chemische Natur des Thromboplastins aus menschlicher Hirnsubstanz. Thrombos. Diathes. haemorrh. (Stuttg) 18: 223 1967;
- 15 Hecht E, Slotta K. H. The chemical nature of the lipid activator in blood coagulation. Amer. J. clin. Path 37: 126 1962;
- 16 Hecht E, Wijngaards G. Recent investigations on the chemical nature of thromboplastin from human brain. Thrombos. Diathes. haemorrh. (Stuttg) 21: 534 1969;
- 17 Hecht E, 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 193: 584 1958;
- 18 Hvatum M, Prydz H. Studies on tissue thromboplastin I. Solubilization with sodium deoxy- cholate. Biochim. biophys. Acta (Amst) 130: 92 1966;
- 19 Irsigler K, Lechner K, Deutsch E, Lomoschitz H. Studies on tissue thromboplastin. II. Species specificity. Thrombos. Diathes. haemorrh. (Stuttg) 14: 18 1965;
- 20 Kazal L. A. Interactions of phospholipids with lipoproteins, with serum and its proteins, and with proteolytic and nonproteolytic enzymes in blood clotting. Transact. N. Y. Acad. Sci. Ser. II 27: 613 1965;
- 21 Kazal L. A. Personal communication by letter of May 20 1966
- 22 Kazal L. A, Miller O. P, Sparks ChE. The properties of complexes of pepsin and brain phospholipid active in thromboplastin generation. Fed. Proc 25: 255 1966;
- 23 Kuhn R, Klesse P. Zur chemischen Konstitution des Lipoids der Thrombokinase. Naturwissenschaften 44: 352 1957;
- 24 Mann F. D, Hum M. Species specificity of thromboplastin. Role of the cothromboplastin reaction. Proc. Soc. exp. Biol. (N. Y.) 79: 19 1952;
- 25 Mills G. A. Chemical nature of tissue coagulins. J. biol. Chem 46: 135 1921;
- 26 Nemerson Y. The phospholipid requirement of tissue factor in blood coagulation. J. clin. Invest 47: 72 1968;
- 27 Quick A. J. The Hemorrhagic Diseases and the Physiology of Hemostasis. 312 Charles C. Thomas; Springfield, Ill., U.S.A: 1942
- 28 Slotta K. H, Deutsch E. Thromboplastic activity of split products of phosphatidyl ethanolamine. Thrombos. Diathes. haemorrh. (Stuttg) 04: 283 1960;
- 29 Soldati B. Untersuchungen über Thrombokinase — Ihre Bedeutung für die Prothrombinzeitbestimmung. Thesis, Zürich: 1941
- 30 Studer A. Contribution à l’étude de la thrombokinase. In Festschrift Emil Bareil. 229 Fa. Hoffrnann-La Roche & Co; Basel, Switserland: 1946
- 31 Thies H. A. Menschliche und tierische Gewebsthrombokinasen. Ihre Eigenschaften und ihre Bedeutung für die Anwendung von Antikoagulantien. Georg Thieme; Stuttgart: 1947
- 32 Wöhlisch E. Die Physiologie und Pathologie der Blutgerinnung. Ergeb. Physiol 28: 443 1929;
- 33 Wooldridge L. C. On intravascular clotting. In: On the Chemistry of the Blood and Other Scientific Papers. 135 Kegan Paul, Trench, Trübner & Co., Ltd.; London, England: 1893
- 34 Wuthier R. E. Purification of lipids from nonlipid contaminants on Sephadex bead columns. J. Lip. Res.. 7/558 1966
- 35 Zahler P. H, Hoelzl Wallach D. F. Isolation of lipid-free plasma membrane proteins by gel filtration on Sephadex LH-20 using 2-chloroethanol-water as solvent. Biochim. biophys. Acta (Amst) 135: 371 1967;
- 36 Zunz E, György P. Contribution à l’étude de l’action des acides aminés, des peptides et des protéoses sur la coagulation du sang. Arch. Int. Physiol 14: 312 1914;
- 37 Zwaal R. F. A, van Deenen L. L. M. Protein patterns of red cel membranes from different mammalian species. Biochim. biophys. Acta (Amst.) 163: 44 1968;