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Thromb Haemost 1977; 38(03): 0630-0639
DOI: 10.1055/s-0038-1651878
Original Article
Schattauer GmbH

Exterior and Interior Events in Early Stage of Thrombin-induced Platelet Aggregation

Shuichi Hashimoto
1   Radioisotope Research Laboratory and Department of Physiological Chemistry, School of Pharmaceutical Sciences, Kitasato University, Tokyo 108, Japan
,
Sachiko Shibata
1   Radioisotope Research Laboratory and Department of Physiological Chemistry, School of Pharmaceutical Sciences, Kitasato University, Tokyo 108, Japan
,
Bonro Kobayashi
1   Radioisotope Research Laboratory and Department of Physiological Chemistry, School of Pharmaceutical Sciences, Kitasato University, Tokyo 108, Japan
› Author Affiliations
Further Information

Publication History

Received 21 July 1976

Accepted 20 April 1977

Publication Date:
04 July 2018 (online)

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Summary

Treatment of washed rabbit platelets with 1 u/ml of thrombin at 37° C resulted in a disappearance from platelets of a protein with 250,000 dalton molecular weight which was shown to be originated from plasma membrane. Parallel loss of adenyl cyclase was noted, and both reactions were complete within 30 sec. From the patterns of disc electrophoretograms, the importance of quick suppression of thrombin action in demonstrating the primary event was stressed.

Thrombin induced an apparent activation of membrane bound phosphodiesterase. This reaction was also complete within 30 sec. The cellular component which contained the enzyme activity was distinct from plasma membrane. Soluble phosphodiesterase was not influenced by thrombin at all.

These reactions required intact platelet cells to react with thrombin, and no reaction was detected when subcellular preparations were treated with thrombin.

Possibility of collaboration of changes in externally located synthetic enzyme with those in internally located degrading enzyme in the early phase of thrombin action on platelets was suggested.

 

  • References

  • 1 Amer M. S, Mayol R. F. 1973; Studies with phosphodiesterase. III. Two forms of the enzyme from human blood platelets. Biochimica et Biophysica Acta 309: 149.
    CrossrefPubMedGoogle Scholar
  • 2 Baenziger N. L, Brodie G. N, Majerus P. W. 1971; A thrombin-sensitive protein of human platelet membranes. Proceedings of the National Academy of Science, USA 68: 240.
    CrossrefPubMedGoogle Scholar
  • 3 Brodie G. N, Baenziger N. L, Chase L. R, Majerus P. W. 1972; The effect of thrombin on adenyl cyclase activity and a membrane protein from human platelets. Journal of Clinical Investigation 51: 81.
    CrossrefPubMedGoogle Scholar
  • 4 Chen L. B, Buchanan J. M. 1975; Plasminogen-independent fibrinolysis by proteases produced by transformed chick embryo fibroblasts. Proceedings of the National Academy of Science, USA 72: 1132.
    CrossrefPubMedGoogle Scholar
  • 5 Cohen I, Bohak Z, de Vries A, Katchalski E. 1969; Thrombosthenin M. Purification and interaction with thrombin. European Journal of Biochemistry 10: 388.
    CrossrefPubMedGoogle Scholar
  • 6 Davey M. G, Lüscher E. F. 1968; Release reactions of human platelets induced by thrombin and other agents. Biochimica et Biophysica Acta 165: 490.
    CrossrefPubMedGoogle Scholar
  • 7 Fairbanks G, Steck T. L, Wallach D. F. H. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10: 2606.
    CrossrefPubMedGoogle Scholar
  • 8 Ganguly P. 1969; Studies on human platelet proteins. II. Effect of thrombin. Blood 33: 590.
    PubMedGoogle Scholar
  • 9 Ganguly P. 1975; Interaction of thrombin with human platelets. British Journal of Haematology 29: 617.
    CrossrefPubMedGoogle Scholar
  • 10 Grette K. 1962 Studies on the mechanism of thrombin-catalyzed hemostatic reactions in blood platelets. Acta Physiologica Scandinavica, Supplementum 195..
    PubMedGoogle Scholar
  • 11 Hagen I. 1975; Effects of thrombin on washed, human platelets: Changes in the subcellular fractions. Biochimica et Biophysica Acta 392: 242.
    CrossrefPubMedGoogle Scholar
  • 12 Hashimoto S, Shibata S, Kobayashi B. 1975; Dependence of platelet adenyl cyclase system on oxidative phosphorylation. Thrombosis et Diathesis Haemorrhagica 34: 42.
    PubMedGoogle Scholar
  • 13 Haslam R. J. 1975. Role of cyclic nucleotides in platelet function. In: Biochemistry and Pharmacology of Platelets. Ciba Foundation Symposium 35 (new series). Elsevier, Excerpta Medica; North-Holland: 121.
    Google Scholar
  • 14 Hynes R. O. 1973; Alteration of cell-surface proteins by viral transformation and by proteolysis. Proceedings of the National Academy of Science, USA 70: 3170.
    CrossrefPubMedGoogle Scholar
  • 15 Kaulen H. D, Gross R. 1971; The differentiation of acid phosphatases of human blood platelets. Thrombosis et Diathesis Haemorrhagica 26: 353.
    PubMedGoogle Scholar
  • 16 Kolassa N, Roos H, Pfleger K. 1972; A separation of purine derivatives by thin-layer chromatography on silica gel plates suitable for metabolic studies. Journal of Chromatography 66: 175.
    CrossrefPubMedGoogle Scholar
  • 17 Krishna G, Weiss B, Brodie B. B. 1968; A simple, sensitive method for the assay of adenyl cyclase. Journal of Pharmacology and Experimental Therapeutics 163: 379.
    PubMedGoogle Scholar
  • 18 Majerus P. W, Brodie G. N. 1972; The binding of phytohemagglutinins to human platelet plasma membranes. Journal of Biological Chemistry 247: 4253.
    PubMedGoogle Scholar
  • 19 Marcus A. J, Zucker-Franklin D, Safier L. B, Ullman H. L. 1966; Studies on human platelet granules and membranes. Journal of Clinical Investigation 45: 14.
    CrossrefPubMedGoogle Scholar
  • 20 Phillips D. R, Agin P. P. 1974; Thrombin substrates and the proteolytic site of thrombin action on human-platelet plasma membranes. Biochimica et Biophysica Acta 352: 218.
    CrossrefPubMedGoogle Scholar
  • 21 Rigas D. A, Osgood E. E. 1955; Purification and properties of the phytohemagglutinin from Phaseolus vulgaris. Journal of Biological Chemistry 212: 607.
    PubMedGoogle Scholar
  • 22 Russell T. R, Terasaki W. L, Appleman M. M. 1973; Separate phosphodiesterases for the hydrolysis of cyclic adenosine 3′, 5′-monophosphate and cyclic guanosine 3′, 5′-monophosphate in rat liver. Journal of Biological Chemistry 248: 1334.
    PubMedGoogle Scholar
  • 23 Salzman E. W, Weisenberger H. 1972. Role of cyclic AMP in platelet function. In: Greengard P, Robison G. A. (eds.) Advances in Cyclic Nucleotide Research. Volume 1: Physiology and Pharmacology of Cyclic AMP. Raven Press; New York: 231.
    Google Scholar
  • 24 Segrest J. P, Jackson R. L. 1972. Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. In: Ginsburg V. (ed.) Method in Enzymology. Volume 28: Complex Carbohydrates, Part B. Academic Press; New York: 54.
    Google Scholar
  • 25 Song S. Y, Cheung W. Y. 1971; Cyclic 3′, 5′-nucleotide phosphodiesterase properties of the enzyme of human blood platelets. Biochimica et Biophysica Acta 242: 593.
    CrossrefPubMedGoogle Scholar
  • 26 Weber T. H. 1969 Isolation and characterization of a lymphocyte-stimulating leukoagglutinin from red kidney beans. Scandinavian Journal of Clinical & Laboratory Investigation, Supplementum 111..
    PubMedGoogle Scholar
  • 27 Weber E, Walter E, Morgenstern E, Mondt H, Rose U, Towliati H, Knaudt E. 1970; Untersuchungen an fraktionierten Plättchenhomogenaten-1. Zur Charakterisierung dreier lysosomaler Enzyme und über die Differenzierung des alpha-Glanulomer. Biochemical Pharmacology 19: 1893.
    CrossrefPubMedGoogle Scholar
  • 28 Weltman J. K, Arnold J. H. 1969; The interaction of a hydrophobic probe with human blood cells. Proceedings of the Society for Experimental Biology and Medicine 131: 546.
    PubMedGoogle Scholar