The Effect of Phospholipase C on Platelet Factor 3 in Human Blood Platelets

Summary

Intact human platelets isolated by gel filtration have been treated with purified phospholipase C. The effect of the enzyme on available and total platelet factor 3 has been tested.

The available procoagulant platelet factor 3 was very low. A further small reduction was observed after incubation with phospholipase C when the enzyme was washed away before testing.

External attack on platelets by phospholipase C led to a marked inactivation of total platelet factor 3.

• References

• 1 Aster R.H, Jandl J.M. Platelet sequestration in man. I. Methods. Journal of clinical Investigation 1964; 43: 843
• 2 Broekman M.J, Westmoreland N.P, Cohen P. An improved method for isolating alpha granules and mitochondria from human platelets. Journal of Cell Biology 1974; 60: 506
• 3 Broekman M.J, Handin R.I, Derksen A, Cohen P. Distribution of phospholipids, fatty acids, and platelet factor 3 activity among subcellular fractions of human platelets. Blood 1976; 47: 963
• 4 Crawford N. Platelet fragments: A microparticulate fraction of platelet-free plasma with adenosine triphosphatase activity. Biochemical Journal 1967; 105: 21 P
• 5 Day H.J, Stormorken H, Holmsen H. Subcellular localization of platelet factor 3 and platelet factor 4. Scandinavian Journal of Haematology 1973; 10: 254
• 6 Falcao L, Gautier A, Lombardi L, Jean G, Probst M. Analogies entre deux types d’organites. Journal of Microscopy 1964; 3: 519
• 7 Gautheron P, Dumont E, Renaud S. Clotting activity of platelet phospholipids in rat and man. Thrombosis et Diathesis Haemorrhagica 1974; 32: 382
• 8 Gollub S, Feldman D, Schecter D.C, Kaplan F.E, Meranze D.R. Thromboplastinase. A new enzyme which destroys thromboplastin. Proceedings of the Society for experimental Biology (N. Y.) 1953; 83: 858
• 9 Husom O. A one stage method for the assay of platelet factor 3. Scandinavian Journal of clinical and laboratory Investigation 1961; 13: 609
• 10 Little C, Aurebekk B, Otnaess A.-B. Purification by affinity chromatography of phospholipase C from Bacillus cereus. FEBS Letters 1975; 52: 175
• 11 Lowry O.H, Rosebrough N.J, Farr A.L, Randall R.J. Protein measurements with the folin phenol reagent. Journal of biological Chemistry 1951; 193: 265
• 12 Maynard J.R, Heckman C.A, Pitlick F.A, Nemerson Y. Association of tissue factor activity with the surface of cultured cells. Journal of clinical Investigation 1975; 55: 814
• 13 Marcus A.J, Zucker-Franklin D, Safier J.B, Ullman H.J. Studies on human platelet granules and membranes. Journal of clinical Investigation 1966; 45: 14
• 14 Otnaess A.-B, Berre Å, Bjørklid E, Prydz H. Phospholipase C from Bacillus cereus and its use in studies of tissue thromboplastin. European Journal of Biochemistry 1972; 27: 238
• 15 Otnaess A.-B, Holm T. The effect of phospholipase C on human blood platelets. Journal of clinical Investigation 1976; 57: 1419
• 16 Otnaess A.-B, Little C, Prydz H. The synthesis of phospholipase C by Bacillus cereus. II. A screening method for mutants. Acta pathologica et microbiologica Scandinavica, Section B 1974; 82: 354
• 17 Otnaess A-B, Little C, Sletten K, Wallin R, Johnsen S, Flengsrud R. Some characteristics of phospholipase C from Bacillus cereus — In preparation.
• 18 Schulz H, Hiepler E. Über die Lokalisierung von gerinnungsphysiologischer Aktivitäten in submikroskopischen Strukturen der Thrombocyten. Klinische Wochenschrift 1959; 37: 273
• 19 Siegel A.M, Lüscher E.F. Non-identity of the α-granules of human blood platelets with typical lysosomes. Nature (London) 1967; 275: 745
• 20 Tangen O, Berman H.J. Gel filtration of blood platelets: A methodological report. In Mannucci P.M, Gorini S. (eds) Platelet function and thrombosis 1972. Plenum Press; New York/London: 235-243
• 21 Vecchione J, Zucker M.B. Procoagulant activity of platelets in recalcified plasma. British Journal of Haematology 1975; 31: 423
• 22 Walsh P.N. Platelet coagulant activities and hemostasis: A hypothesis. Blood 1974; 43: 597
• 23 Zacharski L.R, Mcintyrne O.R. Membrane-mediated synthesis of tissue factor (thromboplastin) in cultured fibroblasts. Blood 1973; 41: 679
• 24 Zwaal R.F.A, Roelofsen B, Comfurius P, Van Deenen L.L.M. Purification and some properties of phospholipase C from Bacillus cereus. Biochimica et Biophysica Acta 1971; 233: 474