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.
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.
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.
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.
5
Cohen I,
Bohak Z,
de Vries A,
Katchalski E.
1969; Thrombosthenin M. Purification and interaction with thrombin. European Journal of Biochemistry 10: 388.
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.
10
Grette K.
1962 Studies on the mechanism of thrombin-catalyzed hemostatic reactions in blood platelets. Acta Physiologica Scandinavica, Supplementum 195..
12
Hashimoto S,
Shibata S,
Kobayashi B.
1975; Dependence of platelet adenyl cyclase system on oxidative phosphorylation. Thrombosis et Diathesis Haemorrhagica 34: 42.
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.
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.
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.
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.
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.
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.
21
Rigas D. A,
Osgood E. E.
1955; Purification and properties of the phytohemagglutinin from Phaseolus vulgaris. Journal of Biological Chemistry 212: 607.
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.
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.
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.
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.
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..
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.
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.
