Thromb Haemost 1983; 49(03): 196-198
DOI: 10.1055/s-0038-1657361
Original Article
Schattauer GmbH Stuttgart

Factors Influencing the Deaggregation of Chymotrypsin-Treated Human Platelets Aggregated by Fibrinogen

R L Kinlough-Rathbone
The Department of Pathology, McMaster University, Hamilton, Canada and the Department of Biochemistry, University of Toronto, Toronto, Canada
,
J F Mustard
The Department of Pathology, McMaster University, Hamilton, Canada and the Department of Biochemistry, University of Toronto, Toronto, Canada
,
M A Packham
The Department of Pathology, McMaster University, Hamilton, Canada and the Department of Biochemistry, University of Toronto, Toronto, Canada
,
E J Harfenist
The Department of Pathology, McMaster University, Hamilton, Canada and the Department of Biochemistry, University of Toronto, Toronto, Canada
› Author Affiliations
Further Information

Publication History

Received 09 March 1983

Accepted 24 March 1983

Publication Date:
18 July 2018 (online)

Summary

Washed human platelets aggregated by ADP can be deaggregated by EDTA or PGE1; provided the release reaction does not occur; when the release reaction occurs platelets deaggregate less readily. Platelets treated with chymotrypsin are aggregated by fibrinogen indicating that fibrinogen receptors may be permanently exposed by this treatment. Fibrinogen-induced aggregation of chymotrypsin-treated platelets provides another method of bringing platelets into close contact with each other. Although EDTA deaggregated chymotrypsin-treated platelets aggregated by fibrinogen in a medium containing a physiological concentration of Ca2+, EDTA did not deaggregate these platelets if they were in a medium without calcium in which the release reaction occurs. In this medium, when ASA was added to prevent the release reaction, EDTA caused deaggregation. More fibrinogen associated with platelets in the medium without calcium than in the calcium-containing medium. In both media, EDTA displaced approximately half of the fibrinogen indicating that deaggregation is not solely dependent on dissociation of fibrinogen from its receptors. Thus when platelets undergo the release reaction, a form of platelet-to-platelet adhesion occurs that is not disrupted by chelation of divalent cations and is therefore not likely to involve only fibrinogen or thrombospondin and fibrinogen since the association of fibrinogen with its receptor requires Ca2+ and the binding of thrombospondin to platelets that have undergone the release reaction is also dependent on Ca2+.

 
  • References

  • 1 Mustard JF, Packham MA, Kinlough-Rathbone RL, Perry DW, Regoeczi E. Fibrinogen and ADP-induced platelet aggregation. Blood 1978; 52: 453-466
  • 2 Marguerie GA, Plow EF, Edgington TS. Human platelets possess an inducible and saturable receptor specific for fibrinogen. J Biol Chem 1979; 254: 5357-5363
  • 3 Bennett JS, Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine. J CUn Invest 1979; 64: 1393-1401
  • 4 Peerschke EI, Zucker MB, Grant RA, Egan JJ, Johnson MM. Correlation between fibrinogen binding to human platelets and platelet aggregability. Blood 1980; 55: 841-847
  • 5 Harfenist EJ, Packham MA, Mustard JF. Reversibility of the association of fibrinogen with rabbit platelets exposed to ADP. Blood 1980; 56: 189-198
  • 6 Marguerie GA, Plow EF, Edgington TS. Interaction of fibrinogen with its platelet receptor as part of a multistep reaction in ADP-induced platelet aggregation. J Biol Chem 1980; 255: 154-161
  • 7 Kornecki E, Niewiarowski S, Morinelli TA, Kloczewiak M. Effects of chymotrypsin and adenosine diphosphate on the exposure of fibrinogen receptors on normal human and Glanzmann’s thrombasthenic platelets. J Biol Chem 1981; 256: 5696-5701
  • 8 Rao GH R, Johnson GJ, White JG. Influence of epinephrine on the aggregation response of aspirin-treated platelets. Prostaglandins Med 1980; 5: 45-18
  • 9 Kinlough-Rathbone RL, Mustard JF, Perry DW, Dejana E, Cazenave J-P, Packham MA, Harfenist EJ. Factors influencing the deaggregation of human and rabbit platelets. Thromb Haemostas 1983; 49: 162-167
  • 10 Mustard JF, Perry DW, Kinlough-Rathbone RL, Packham MA. Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol 1975; 228: 1757-1765
  • 11 Kinlough-Rathbone RL, Mustard JF, Packham MA, Perry DW, Reimers H-J, Cazenave J-P. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
  • 12 Macfarlane DE, Walsh PN, Mills DC B, Holmsen H, Day HJ. The role of thrombin in ADP-induced platelet aggregation and release: a critical evaluation. Br J Haematol 1975; 30: 457-463
  • 13 Lages B, Weiss HJ. Dependence of human platelet functional responses on divalent cations: aggregation and secretion in heparinand hirudin-anticoagulated platelet-rich plasma and the effects of chelating agents. Thromb Haemostas 1981; 45: 173-179
  • 14 Greenberg JP, Packham MA, Guccione MA, Harfenist EJ, Orr JL, Kinlough-Rathbone RL, Perry DW, Mustard JF. Effect of pretreatment of human or rabbit platelets with chymotrypsin on their responses to human fibrinogen and aggregating agents. Blood 1979; 54: 753-765
  • 15 Niewiarowski S, Budzynski AZ, Morinelli TA, Brudzynski TM, Stewart GJ. Exposure of fibrinogen receptor on human platelets by proteolytic enzymes. J Biol Chem 256: 917-925
  • 16 Grant RA, Zucker MB. Effects of chymotrypsin on the release reaction and aggregation of blood platelets. Proc Soc Exp Biol Med 1980; 165: 114-117
  • 17 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
  • 18 Ardlie NG, Perry DW, Packham MA, Mustard JF. Influence of apyrase on the stability of suspensions of washed rabbit platelets. Proc Soc Exp Biol Med 1971; 136: 1021-1023
  • 19 Greenberg J, Packham MA, Cazenave J-P, Reimers H-J, Mustard JF. Effect on platelet function of removal of platelet sialic acid by neuraminidase. Lab Invest 1975; 32: 476-484
  • 20 Lawrie JS, Ross J, Kemp GD. Purification of fibrinogen and the separation of its degradation products in the presence of calcium ions. Biochem Soc Transact 1979; 7: 693-694
  • 21 McFarlane AS. Efficient trace labelling of proteins with iodine. Nature 1958; 182: 53
  • 22 Kinlough-Rathbone RL, Packham MA, Mustard JF. The effect of prostaglandin E1 on platelet function in vitro and in vivo. Br J Haematol 1970; 19: 559-571
  • 23 Davies JA, Essien E, Cazenave J-P, Kinlough-Rathbone RL, Gent M, Mustard JF. The influence of red blood cells on the effects of aspirin or sulfinpyrazone on platelet adherence to damaged rabbit aorta. Br J Haematol 1979; 42: 283-291
  • 24 Gartner TK, Gerrard JM, White JG, Williams DC. Fibrinogen is the receptor for the endogenous lectin of human platelets. Nature 1981; 289: 688-690
  • 25 Phillips DR, Jennings LK, Prasanna HR. Ca2+-mediated association of glycoprotein G (thrombin-sensitive protein, thrombospondin) with human platelets. J Biol Chem 1980; 255: 11629-11632