There is controversy in the literature regarding the effects of plasmin on human platelets. We have studied the effects of plasmin on platelet glycoproteins, aggregation, shape change and secretion and found them to be dependent on experimental conditions: (a) Plasmin's effects on human platelets are only seen in gel-filtered platelets (GFP), presumably because in platelet rich plasma plasmin is bound to a2-antiplasmin; (b) in GFP to which fibrinogen has been added, platelet function remains intact; and (c) in the absence of fibrinogen, the effect of plasmin on GFP depends on whether stirring is performed or not. With stirring, platelets undergo shape change, secretion and aggregation in response to added plasmin. Aggregation is much stronger when CaClz1 mM is added. Without stirring, preincubation of GFP with plasmin leads to inhibition of platelet aggregation induced by subsequent platelet stimuli (thrombin, collagen, ristocetin or U46619). We have demonstrated that plasmin is a true platelet activating agent, in the sense that it induces platelet shape change and secretion. Plasmin will induce aggregation when added to stirred GFP. This may be because stirring protects glycoprotein (GP) IIbIIIa bound fibrinogen from being degradated by plasmin. When added to unstirred GFP, GP IIbIIIa bound fibrinogen may be readily accessible to degradation by plasmin, which may then behave like a platelet inhibitor.